Heterocyclic compounds as inhibitors of beta-lactamases

ABSTRACT

This invention discloses and claims methods for inhibiting bacterial β-lactamases and treating bacterial infections by inhibiting bacterial β-lactamases in man or an animal comprising administering a therapeutically effective amount to said man or said animal of a compound, or pharmaceutically acceptable salt thereof, of formula (I) either alone or in combination with a β-lactamine antibiotic wherein said combination can be administered separately, together or spaced out over time. Pharmaceutical compositions comprising a compound of formula (I), or a combination of a compound of formula (I) and a therapeutically effective amount of a β-lactamine antibiotic, and a pharmaceutically acceptable carrier are also disclosed and claimed.

This application is a continuation of International Application No.PCT/FR03/00243 filed Jan. 27, 2003, which claims the benefit of priorityof French Application No. 02 00951, filed Jan. 28, 2002.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention relates to heterocyclic compounds, endowed withbeta-lactamase inhibitory properties, and therefore are of interest incombating infectious diseases or in the prevention of them, in the formof a combination with various antibiotic compounds of β-lactamine type,in order to reinforce their effectiveness in combating the pathogenicbacteria which produce β-lactamases.

2. Description of the Art

It is well known that the enzymatic inactivation of antibiotics ofβ-lactamine type, whether compounds of penicillin or cephalosporin type,in the treatment of bacterial infections is a obstacle for this type ofcompound. This inactivation consists of a degradation process of theβ-lactamines and constitutes one of the mechanisms by which the bacteriacan become resistant to treatments. It is therefore desirable to actagainst this enzymatic process by combining an agent capable ofinhibiting the enzyme with the antibacterial agent of β-lactamine type.When an inhibitor of β-lactamase is used in combination with anantibiotic of β-lactamine type, it can thus reinforce its effectivenessagainst certain microorganisms.

SUMMARY OF THE INVENTION

The invention therefore relates to the compounds corresponding to theformula (I):

in which:R₁ represents a hydrogen atom, a COOH, CN, COOR, CONR₆, R₇, (CH₂)_(n′)R₅or

radicalR is chosen from group constituted by an alkyl radical containing 1 to 6carbon atoms, optionally substituted by a pyridyl or carbamoyl radical,a —CH₂-alkenyl radical containing a total of 3 to 9 carbon atoms, arylcontaining 6 to 10 carbon atoms or aralkyl containing 7 to 11 carbonatoms, the nucleus of aryl or aralkyl radical being optionallysubstituted by an OH, NH₂, NO₂, alkyl radical containing 1 to 6 carbonatoms, alkoxy containing 1 to 6 carbon atoms or by one or more halogenatoms,R₆ and R₇, identical or different, are chosen from the group constitutedby a hydrogen atom, an alkyl radical containing 1 to 6 carbon atoms,aryl containing 6 to 10 carbon atoms and aralkyl containing 7 to 11carbon atoms, optionally substituted by a carbamoyl, ureido ordimethylamino radical, and an alkyl radical containing 1 to 6 carbonatoms substituted by a pyridyl radical, n′ is equal to 1 or 2 and R₅ ischosen from the group constituted by a COOH, CN, OH, NH₂, CO—NR₆R₇,COOR, OR, OCHO, OCOR, OCOOR, OCONHR, OCONH₂, NHR, NHCOH, NHCOR, NHSO₂R,NH—COOR, NH—CO—NHR or NHCONH₂ radical, R, R₆ and R₇ being as definedabove;R₂ represents a hydrogen atom or a (CH₂)_(n′1)R₅ group, n′₁ being equalto 0, 1 or 2, and R₅ being as defined above;R₃ represents a hydrogen atom or an alkyl radical containing 1 to 6carbon atoms;A represents a bond between the two carbons which carry R₁ and R₂ or a

group, R₄ representing a hydrogen atom or a (CH₂)_(n′1)R₅ group, n′₁ andR₅ being as defined above, the dotted line representing an optionaladditional bond with one or other of the carbons which carry thesubstituents R₁ and R₂,n is equal to 1 or 2,X represents a divalent —C(O)—B— group linked to the nitrogen atom bythe carbon atom, B represents a divalent —O—(CH₂)_(n″)— group linked tothe carbonyl by the oxygen atom, an —NR₈—(CH₂)_(n′)— or —NR₈—O— grouplinked to the carbonyl by the nitrogen atom, n″ is equal to 0 or 1 andR₈, in the case of —NR₈—(CH₂)_(n″)— is chosen from the group constitutedby a hydrogen, an OH, R, OR, Y, OY, Y₁, OY₁, Y₂, OY₂, Y₃,OCH₂CH₂SO_(m)R, OSiR_(a)R_(b)R_(c) and SiR_(a)R_(b)R_(c) radical, and inthe case of —NR₈—O— is chosen from the group constituted by a hydrogen,an R, Y, Y₁, Y₂, Y₃ and SiR_(a)R_(b)R_(c) radical, R_(a), R_(b) andR_(c) representing individually a linear or branched alkyl radicalcontaining 1 to 6 carbon atoms or an aryl radical containing 6 to 10carbon atoms, R being as defined previously and m being equal to 0, 1 or2,Y is chosen from the group constituted by the COH, COR, COOR, CONH₂,CONHR, CONHOH, CONHSO₂R, CH₂COOH, CH₂COOR, CH₂CONHOH, CH₂CONHCN,CH₂tetrazole, protected CH₂tetrazole, CH₂SO₃H, CH₂SO₂R, CH₂PO(OR)₂,CH₂PO(OR)(OH), CH₂PO(R)(OH) and CH₂PO(OH)₂ radicals,Y₁ is chosen from the group constituted by the SO₂R, SO₂NHCOH, SO₂NHCOR,SO₂NHCOOR, SO₂NHCONHR SO₂NHCONH₂ and SO₃H radicals,Y₂ is chosen from the group constituted by the PO(OH)₂, PO(OR)₂,PO(OH)(OR) and PO(OH)(R) radicals,Y₃ is chosen from the group constituted by the following radicals:tetrazole, tetrazole substituted by the R, squarate, NH or NR tetrazole,NH or NR tetrazole substituted by the R radical, NHSO₂R and NRSO₂R, Rbeing as defined above;R₁, R₂ and R₃ not representing all three at the same time a hydrogenatom when n is equal to 1 and A represents a

group in which R4 is a hydrogen atom and

-   -   either X represents the —C(O)—O—(CH₂)_(n″) group in which n″ is        0 or 1,    -   or X represents the —CO—NR₈—(CH₂)_(n″) group in which n″ is 1        and R₈ is the isopropyl group,    -   or X represents the —CO—NR₈—(CH₂)_(n″) group in which n″ is 0        and R₈ is hydrogen or phenyl,        as well as the salts of these compounds with mineral or organic        bases or acids, as well as the internal salts in the form in        which they can, if appropriate, be presented.

The compounds of formula (I) and their salts are described and claimedin the International Patent Application No. PCT/FR01/02418 filed on the24 Jul. 2001, claiming priority from the French Application No.^(o)0010121 filed on the 1 Aug. 2000.

The compounds of formula (I) are presented in the form of pureenantiomers or pure diastereoisomers or in the form of a mixture ofenantiomers in particular of racemates, or mixtures of diastereoisomers.Moreover, the substituents R₁, R₂, R₄ taken individually on the one handand X on the other can be in cis and/or trans position with respect tothe ring on which they are fixed, and the compounds of formula (I) aretherefore presented in the form of cis isomers or trans isomers ormixtures.

By alkyl radical containing 1 to 6 carbon atoms, is meant the methyl,ethyl radical, as well as propyl, butyl, pentyl or hexyl linear,branched or cyclic.

By —CH₂-alkenyl radical containing 3 to 9 carbon atoms, is meant forexample the allyl radical, or a butenyl, pentenyl or hexenyl radical.

By aryl radical containing 6 to 10 carbon atoms, is meant a phenyl ornaphthyl radical.

By aralkyl radical containing 7 to 11 carbon atoms, is meant a benzyl,phenethyl or methylnaphthyl radical.

By alkyloxy radical containing 1 to 6 carbon atoms, is meant inparticular the methoxy, ethoxy, propoxy, isopropoxy radical, as well asbutoxy, isobutoxy, dry-butoxy or tert-butoxy radical.

By halogen atom, is meant a fluorine, chlorine, bromine or iodine atom.

By squarate radical, is meant the radical of formula:

Among the salts with acids of the products of formula (I), there can bementioned amongst others, those formed with mineral acids, such ashydrochloric, hydrobromic, hydroiodic, sulphuric or phosphoric acids orwith organic acids such as formic, acetic, trifluoroacetic, propionic,benzoic, maleic, fumaric, succinic, tartaric, citric, oxalic, glyoxylic,aspartic acid, alkanesulphonics, such methane and ethane sulphonicacids, arylsulphonics such as benzene and paratoluenesulphonic acids.

Among the salts with bases of the products of formula (I), there can bementioned, amongst others, those formed with mineral bases such as, forexample, sodium, potassium, lithium, calcium, magnesium or ammoniumhydroxide or with organic bases such as, for example, methylamine,propylamine, trimethylamine, diethylamine, triethylamine,N,N-dimethylethanolamine, tris(hydroxymethyl)amino methane,ethanolamine, pyridine, picoline, dicyclohexylamine, morpholine,benzylamine, procaine, lysine, arginine, histidine, N-methylglucamine,or also phosphonium salts, such as alkyl-phosphoniums,aryl-phosphoniums, alkyl-aryl-phosphoniums, thealkenyl-aryl-phosphoniums or quaternary ammonium salts such the salt oftetra-n-butyl-ammonium.

A subject of the invention is the use of the compounds of formula (I),as well as their pharmaceutically acceptable salts, for the preparationof a medicament intended to inhibit the production of β-lactamases bypathogenic bacteria.

A subject of the invention is also the use of the compounds of formula(I), as well as their pharmaceutically acceptable salts, which inhibitthe production of β-lactamases by pathogenic bacteria, for thepreparation of a medicament intended for the treatment of bacterialinfections in man or animals.

A particular subject of the invention is a use according to what hasgone before, characterized in that the compounds correspond to formula(I) in which n is equal to 1 and A and R₂ are as defined above, R₃represents a hydrogen atom, R₁ represents a hydrogen atom, a COOR orCONR₆R₇ radical, R₆ and R₇ being as defined above and X represents a—C(O)—B— group in which B represents an —O—(CH₂)_(n″)— or—NR₈—(CH₂)_(n″)— group, n″ being equal to O and R₈ having the values asdefined above and, more particularly the values Y, Y₁ and OY₁, and moreparticularly, from among these, those corresponding to formula (I) inwhich A represents a

group in which R₄ represents a hydrogen atom, R₂ represents a hydrogenatom and B represents an NR₈—(CH₂)_(n″)— group in which n″ is equal to Oand R₈ represents an OY₁ radical.

A quite particular subject of the invention is a use as defined above,characterized in that the compounds are chosen from the list constitutedby:

-   cis-7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-4-propanoic-acid,-   trans diphenylmethyl 7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetate,-   cis diphenylmethyl 7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetate,-   trans phenylmethyl    3-benzoyl-2-oxo-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate,-   trans phenylmethyl    2-oxo-3-(sulphooxy)-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate,-   6-[[(4-methylphenyl)sulphonyl]oxy]-1,6-diazabicyclo[3.2.1]octan-7-one,-   6-[(methylsulphonyl)oxy]-1,6-diazabicyclo[3.2.1]octan-7-one,-   6-[(4-nitrophenyl)sulphonyl]oxy]-1,6-diazabicyclo[3.2.1]octan-7-one,-   trans diphenylmethyl    7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-2-carboxylate,-   trans(4-nitrophenyl)methyl    7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-2-carboxylate,-   trans-7-oxo-6-oxa-1-azabicyclo[3.2.1.]octane-2 carboxylic acid,-   trans phenylmethyl    7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,-   trans phenylmethyl    7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,-   trans phenylmethyl    7-oxo-6-[(phenylsulphonyl)oxy]-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,-   trans phenylmethyl    7-oxo-6-[(2-thienylsulphonyl)oxy]-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,-   trans-6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic    acid,-   trans methyl    6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,-   trans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-7-oxo-N-(phenylmethyl)-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-7-oxo-N-(2-pyridinylmethyl)-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-7-oxo-N-[2-(3-pyridinyl)ethyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-7-oxo-N-[2-(4-pyridinyl)ethyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-7-oxo-N-[2-(2-pyridinyl)ethyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-N-[3-(aminocarbonyl)phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-N-[4-(dimethylamino)phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-N-[3-(dimethylamino)phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-7-oxo-N-[(4-pyridinyl)methyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-7-oxo-N-(3-pyridinylmethyl)-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-N-(1-amino-1-oxo-3-phenyl-2-propyl)-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-N-(2-amino-2-oxoethyl)-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-N-[3-[(aminocarbonyl)amino]phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans-N-(2-amino-2-oxo-1-phenylethyl)-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,-   trans 2-amino-2-oxoethyl    7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,-   trans 2-(4-pyridinyl)ethyl    7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,-   trans 2-(2-pyridinyl)ethyl    7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,-   6-(sulphooxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one,-   3-methoxy-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one, as    well as their salts.

A subject of the invention is also a use as defined above, characterizedin that within a medicament, the compound of formula (I) is combinedwith an antibiotic of β-lactamine type chosen from the group constitutedby the penams, the penems, the carbapenems, the cephems, thecarbacephems, the oxacephems, the cephamycins and the monobactams.

By β-lactamines, is meant for example the penicillins such asamoxicillin, ampicillin, azlocillin, mezlocillin, apalcillin,hetacillin, bacampicillin, carbenicillin, sulbenicillin, ticarcillin,piperacillin, azlocillin, mecillinam, pivmecillinam, methicillin,ciclacillin, talampicillin, aspoxicillin, oxacillin, cloxacillin,dicloxacillin, flucloxacillin, nafcillin or pivampicillin, thecephalosporins such as cephalothin, cephaloridine, cefaclor, cefadroxil,cefamandole, cefazolin, cephalexin, cephradine, ceftizoxime, cefoxitin,cephacetrile, cefotiam, cefotaxime, cefsulodin, cefoperazone,ceftizoxime, cefmenoxime, cefmetazole, cephaloglycin, cefonicid,cefodizime, cefpirome, ceftazidime, ceftriaxone, cefpiramide,cefbuperazone, cefozopran, cefepime, cefoselis, cefluprenam, cefuzonam,cefpimizole, cefclidin, cefixime, ceftibuten, cefdinir, cefpodoximeaxetil, cefpodoxime proxetil, cefteram pivoxil, cefetamet pivoxil,cefcapene pivoxil or cefditoren pivoxil, cefuroxime, cefuroxime axetil,loracarbacef, latamoxef, the carbapenems such as imipenem, meropenem,biapenem or panipenem and the monobactams such as aztreonam andcarumonam, as well as their salts.

The compounds of formula (I) or their pharmaceutically acceptable salts,can be administered at the same time as the dose of an antibiotic of theβ-lactamine type, or separately, preferably after it. This can becarried out in the form of a mixture of the two active ingredients or inthe form of a pharmaceutical combination of the two separate activeingredients.

The dose of the compounds of formula (I) and of their pharmaceuticallyacceptable salts can of course vary within wide limits and mustnaturally be adapted, in each particular case, to the individualconditions and to the pathogenic agent, which produces β-lactamase, tobe combated. In general, a daily dose can range from 0.1 toapproximately 10 g as may be convenient.

Moreover, the ratio of the β-lactamase inhibitor of formula (I) or ofthe pharmaceutically acceptable salt of the latter to the antibiotic ofβ-lactamine type can also varies vary within wide limits and must beadapted, in each particular case, to the individual conditions. Ingeneral, a ratio ranging from approximately 1:20 to approximately 1:1must be indicated.

The medicaments as defined above are employed in the form ofpharmaceutical compositions in a mixture with an organic or mineral,inert pharmaceutical excipient, adapted to the sought administrationmethod, and a subject of the invention is also said pharmaceuticalcompositions.

These compositions can be solid or liquid and be presented in thepharmaceutical forms commonly used in human medicine, such as forexample, plain or sugar-coated tablets, gelatin capsules, granules,suppositories, injectable preparations, ointments, creams, gels, theyare prepared according to the usual methods. The active ingredient oringredients can be incorporated with excipients usually employed inthese pharmaceutical compositions, such as talc, gum arabic, lactose,starch, magnesium stearate, cocoa butter, aqueous or non-aqueousvehicles, fatty substances of animal or vegetable origin, paraffinderivatives, glycols, various wetting, dispersing or emulsifying agents,preservatives.

These compositions can also be presented in the form of a lyophilisateintended to be dissolved extemporaneously in an appropriate vehicle forexample apyrogenic sterile water.

The compounds of formula (I) can be prepared by a process comprising:

a) a stage during which a carbonylation agent, if appropriate in thepresence of a base, is reacted with a compound of formula (II):

in which:R′₁ represents a hydrogen atom or a CN, protected COOH, COOR′,(CH₂)n′R′₅, CONR₆R₇ or protected

radical;n′, R₆ and R₇ having the above definitions and R′ and R′₅ having theabove definitions of R and R₅ respectively, in which the reactivefunctions which are optionally present are protected;R′₂ represents a hydrogen atom or a (CH₂)n′₁R′₅ group, n′₁ and R′₅ beingas defined above;R₃ is as defined previously;A′ represents a bond between the two carbons which carry R′₁ and R′₂ ora

group, R′₄ representing a hydrogen atom or a (CH₂)n′₁R′₅ group, n′₁ andR′₅ being as defined above, the dotted line representing an optionalbond with one or other of the carbons which carry substituents R′₁ andR′₂;n is as defined previously;HZ represents a HO—(CH₂)n″—, HNR′₈—(CH₂)_(n″)— or HNR′₈—O— group, n″being as defined previously and R′₈ representing a hydrogen atom, aprotected OH, R′, OR′ radical, a Y′ or OY′ radical, Y′ being chosen fromthe COH, COR′, COOR′, CONH₂, CONHR′, protected CONHOH, CONHSO₂R′,protected CH₂COOH, CH₂COOR′, protected CH₂CONHOH, CH₂CONHCN,CH₂tetrazole substituted by R′, CH₂SO₂R′, CH₂PO(OR′)₂, protected CH₂SO₃,protected CH₂PO(OR′)OH, protected CH₂PO(R′)OH, protected CH₂PO(OH)₂groups, a Y′₁ or OY₁ radical, Y′₁ being chosen from the SO₂R′, SO₂NHCOH,SO₂NHCOR′, SO₂NHCOOR′, SO₂NHCONH₂, SO₂NHCONHR′ and protected SO₃Hgroups, a Y′₂ or OY′₂ radical, Y′₂ representing a protected PO(OH)₂,protected PO(OH)(OR′), protected PO(OH)R′ or PO(OR′)₂ group, or a Y′₃radical, Y′₃ being chosen from the protected tetrazole, tetrazolesubstituted by the R′ radical, protected NH or NR′ tetrazole, NH or NR′tetrazole substituted by the R′ radical, NHSO₂R′ and NR′SO₂R′ groups, R′being as defined above;with a view to obtaining an intermediate compound of formula:

in which:R′₁, R′₂, R₃, A′ and n have the same meanings as above and either X₁ isa hydrogen atom and X₂ represents a -Z-CO—X₃ group, X₃ representing theremainder of the carbonylation agent, or X₂ is a -ZH group and X₁represents a CO—X₃ group, X₃ being as defined previously;b) a stage during which the intermediate obtained previously iscyclized, in the presence of a base; and in that:c) if appropriate, Stage a) is preceded and/or Stage b) is followed byone or more of the following reactions, in an appropriate order:

-   -   protection of the reaction functions,    -   deprotection of the reaction functions,    -   esterification,    -   saponification,    -   sulphation,    -   phosphation    -   amidification,    -   acylation,    -   sulphonylation;    -   alkylation;    -   introduction of a double bond;    -   formation of a urea group;    -   introduction of a tetrazole group;    -   reduction of the carboxylic acids;    -   dehydration of amide to nitrile;    -   salification;    -   ion exchange;    -   resolution or separation of diastereoisomers;    -   oxidation of sulphide to sulphoxide and/or sulphone.

As a carbonylation agent, a reagent can be employed such as phosgene,diphosgene, triphosgene, an aryl chloroformate such as phenyl orp-nitrophenyl chloroformate, an aralkyl chloroformate such as benzylchloroformate, an alkyl or alkenyl chloroformate such as methyl or allylchloroformate, an alkyl dicarbonate such as tert-butyl dicarbonate,carbonyl-diimidazole and their mixtures.

The reaction preferably takes place in the presence of a base or amixture of bases which neutralize the acid formed. It can in particularbe an amine such as triethylamine, diisopropylethylamine, pyridine,dimethylaminopyridine. However, the operation can also be carried outusing the starting product of formula II as base. It is then used inexcess. An illustration is given in the experimental part.

If appropriate, the product of formula II is employed in the form of anacid salt, for example a hydrochloride or a trifluoroacetate.

As base in Stage b), amines, or also hydrides, alcoholates, amides orcarbonates of alkali or alkaline-earth metals can also be used.

The amines can be chosen for example from the above list.

As hydride, sodium or potassium hydride can in particular be used.

As alkali metal alcoholate, potassium t-butylate is preferably used.

As alkali metal amide lithium bis(trimethylsilyl) amide can inparticular be used.

As a carbonate, sodium or potassium carbonate or bicarbonate can inparticular be used.

If appropriate, the intermediate of formula III can be obtained in theform of an acid salt generated during the carbonylation reaction and inparticular a hydrochloride. It is then employed in the cyclizationreaction in this form.

If appropriate, the cyclization can be carried out without isolation ofthe intermediate of formula III.

The reactions mentioned in Stage c) are generally standard reactions,which are well-known to a person skilled in the art.

The reactive functions which it is convenient and appropriate to protectare carboxylic acid, amine, amide, hydroxy and hydroxylamine functions.

The protection of the acid function is in particular carried out in theform of alkyl esters, allyl, benzyl, benzhydryl or p-nitrobenzyl esters.

Deprotection is carried out by saponification, acid hydrolysis,hydrogenolysis, or also cleavage using soluble complexes of palladium 0.

Examples of these protections and deprotections are provided hereafterin the experimental part.

Protection of the amines and amides is in particular carried out in theform of benzylated derivatives, in the form of carbamates, in particularof allyl, benzyl, phenyl or tertbutyl, or also in the form of silylatedderivatives such as tertbutyl dimethyl, trimethyl, triphenyl or alsodiphenyl tertbutyl-silyl derivatives.

Deprotection is carried out, according to the nature of the protectivegroup, by sodium or lithium in liquid ammonia, by hydrogenolysis orusing soluble complexes of palladium 0, by the action of an acid, or bythe action of tetrabutylammonium fluoride.

Examples are provided hereafter in the experimental part.

The protection of hydroxylamines is carried out in particular in theform of benzyl or allyl ethers.

Cleavage of the ethers is carried out by hydrogenolysis or by usingsoluble complexes of palladium 0.

A illustration is provided below in the experimental part.

Protection of the alcohols is carried out in a standard fashion, in theform of ethers, esters or carbonates. The ethers can be alkyl oralkoxyalkyl ethers, preferably methyl or methoxyethoxymethyl ethers,aryl or preferably aralkyl ethers, for example benzyl, or silylatedethers, for example the silylated derivatives mentioned above. Theesters can be any cleavable ester known to a person skilled in the artand preferably the acetate, propionate or benzoate or p-nitrobenzoate.The carbonates can be, for example, methyl, tertbutyl, allyl, benzyl orp-nitrobenzyl carbonates.

Deprotection is carried out by means known to a person skilled in theart, in particular, saponification, hydrogenolysis, cleavage by solublecomplexes of palladium 0, hydrolysis in acid medium or also, for thesilylated derivatives, treatment with tetrabutylammonium fluoride.

Examples are provided in the experimental part.

The sulphation reaction is carried out by the action of SO₃-aminecomplexes such as SO₃-pyridine or SO₃-dimethylformamide, operating inpyridine, the salt formed, for example the pyridine salt, can then beexchanged for example by a salt of another amine, of a quaternaryammonium or of an alkali metal. Examples are provided in theexperimental part.

The phosphation reaction is carried out for example by the action of achlorophosphate such as dimethyl, dibenzyl or diphenyl chlorophosphate.

The amidification reaction is carried out starting from the carboxylicacid using an activation agent such as an alkyl chloroformate or EDCI,by the action of ammonium hydroxide or of an appropriate amine or theiracid salts. Examples are provided hereafter in the experimental part.

The acylation and sulphonylation reactions are carried out on thehydroxyureas by the action respectively of a halide or an anhydride ofan appropriate carboxylic acid or a halide of an appropriate sulphonicacid. Several examples are provided hereafter in the experimental part.

The alkylation reaction is carried out by the action on the hydroxylatedderivatives of an alkyl halide or an alkyl halide substituted, inparticular, by a free or esterified carboxy radical. Illustrations areprovided hereafter in the experimental part.

The final optional introduction of a double bond, which is thenpreferably situated between the carbon atoms which carry R₄ and R₁, iscarried out by the action of a halogenated derivative of selenium thenoxidation, according to the methods known to a person skilled in theart. An example is shown hereafter in the experimental part.

Formation of a urea group, which relates to the substituent R₈ ispreferably carried out by the action of an appropriate isocyanate on thefree NH. An example is shown hereafter in the experimental part.

Introduction of a tetrazole group is carried out by the action of ahalogenated derivative, preferably fluorinated, of the protected orsubstituted tetrazole. Deprotection can be carried out byhydrogenolysis.

Reduction of the acids to alcohols can be carried out by the action of aborane or via a mixed anhydride intermediate, by the action of analkaline borohydide. The mixed anhydride is prepared for example usingan alkyl chloroformate. An illustration of this is provided in theexperimental part.

Dehydration of the amide to a nitrile can occur under the carbonylationand cyclization reaction conditions.

Oxidation of the sulphide to sulphoxide and/or sulphone can be carriedout by the action of a peracid such as metachloroperbenzoic orperphthalic acid or any other reagent known to a person skilled in theart.

Salification by acids is, if appropriate, carried out by adding an acidin soluble phase to the compound. Salification by bases can concerneither the compounds comprising an acid function, in particular carboxy,or those comprising a sulphooxy function or a phosphoric acid derivativeor those comprising a heterocycle with an acid character. In the firstcase, the operation is carried out by adding an appropriate base such asthose mentioned previously. In the second case, the pyridinium salt isobtained directly during the action of the SO₃-pyridine complex and theother salts are obtained from this pyridinium salt. In one or othercase, the operation can also be carried out by ion exchange on resin.Examples of salifications are shown hereafter in the experimental part.

Separation of the enantiomers and diastereoisomers can be carried outaccording to the techniques known to a person skilled in the art, inparticular chromatography.

In addition to the processes described previously, the compounds offormula (I) can of course be obtained by methods which use at the starta compound of formula (II) in which R′₁, A′, R′₂, R₃ and HZ have thevalues which lead directly (without conversion) to those of thecompounds that one wishes to prepare. If appropriate, those of thesevalues which would contain reaction functions as mentioned above arethen protected, deprotection occurring at the end of the cyclizationstage b or at any other opportune moment in the synthesis. Theprotections and deprotections are then carried out as described above.

Such methods are provided hereafter in the experimental part.

The products of formula (II) are known or can be prepared according tomethods known to a person skilled in the art. References from theliterature as well as the preparations are provided hereafter in theexperimental part.

EXAMPLES

The following examples illustrate the invention, without howeverlimiting the scope.

In the examples which follow the following abbreviations have been used:

DEAD: diethyl azo-dicarboxylate

TEA: triethylamine

DMAP: 4-dimethylamino-pyridine

EDCI: 1-(3-dimethylamino-propyl)-3-ethylcarbo-diimide hydrochloride

THF: tetrahydrofuran

AIBN: 2.2′-azo-bis-isobutyronitrile

M: molar molecular mass

MS: mass spectrometry

EI: electron impact

SIMS: secondary ion mass spectrometry

FAB: fast atom bombardment

Example 1 cis diphenylmethyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-4-propanoate

3.16 g (10.6 mmoles) of the hydrochloride of3-oxo-1-(phenylmethyl)-4-piperidinepropanoic acid (M=297.7 g) (describedin the Japanese Patent Application J54098-772) is mixed with 100 ml ofethanol and the reaction medium is cooled down to 10° C. 1.84 g of NaBH₄is added over 15 minutes, under a stream of nitrogen, whilst maintainingthe temperature between 8 and 13° C. The temperature is allowed to riseto ambient temperature and left in contact for 1 hour 30 minutes.Another 380 mg of NaBH₄ is added and the reaction medium is leftovernight at ambient temperature.

The solvent is evaporated off under reduced pressure, followed by takingup in 50 ml of water and adjusting the pH from 10 to 2 usingconcentrated hydrochloric acid. Evaporation is again carried out underreduced pressure. The solid residue (approximately 10.8 g) is washedtwice with 100 ml of ethanol then the solvent is evaporated off underreduced pressure.

In this way 3.10 g of the hydrochloride of3-hydroxy-1-(phenylmethyl)-4-piperidinepropanoic acid (M=299.7 g) isobtained, which corresponds to a yield of 97%.

3.10 g (10.3 mmoles) of the compound obtained previously is diluted in100 ml of ethanol then 900 mg of 10% Pd/C by prehydrogenated weight andin 30 ml of ethanol is added to it.

The reaction medium is left under a hydrogen atmosphere at normalpressure overnight, then the catalyst is eliminated by filtration andthe ethanol by evaporation under reduced pressure.

1.90 g of the hydrochloride of trans-3-hydroxy-4-piperidinepropanoicacid (M=209.6 g), is obtained i.e. a yield of 88%.

1.79 g (8.54 mmoles) of the compound obtained previously is mixed with20 ml of ethanol and 20 ml of water.

Then concentrated soda is added until the pH is approximately 8.5.

Then, 1 ml of allyl chloroformate and concentrated soda are added so asto maintain the pH between 8 and 9.

The reaction mixture is extracted with ethyl acetate then the aqueousphase is acidified to pH 2 by adding concentrated hydrochloric acid andreextracting with ethyl acetate. After drying and evaporating thesolvent under reduced pressure, 1.69 g of crude product is obtainedwhich is taken up in a mixture of dichloromethane and ethanol, followedby filtering and again evaporating the solvent under reduced pressure.

In this way 1.40 g oftrans-3-hydroxy-1-[(2-propenyloxy)carbonyl]-4-piperidinepropanoic acid(M=257 g) is obtained, i.e. a yield of 60%.

3.24 g (12.6 mmoles) of the above hydroxy-acid and 6.4 g oftriphenylphosphine are dissolved in 60 ml of THF at 0° C. under anitrogen atmosphere. Then 2.5 ml of DEAD is added and after 15 minutesthe reaction mixture is evaporated under reduced pressure in order toobtain 12 g of crude product. Purification is carried out bychromatography on silica eluting progressively with a mixture ofdichloromethane and ethyl acetate 9/1, 8/2, 7/3 in order to separate thecis and trans lactones.

In this way 2.72 g of cis lactone is obtained in a mixture with reducedDEAD and phosphine oxide.

This product is resolubilized in 10 ml of DME and 8 ml of a 1N solutionof NaOH is added. After contact for 1 hour, the reaction mixture isextracted twice with ethyl acetate, then acidified to pH 2 with 2N HCl,and reextracted with ethyl acetate. After drying and evaporating thesolvent under reduced pressure, 1.07 g of hydroxy-acid is obtained.

1.0 g of crude hydroxy-acid is dissolved in a mixture of 5 ml ofdichloromethane and 2 ml of methanol, then treated with an excess ofdiphenyldiazomethane in dichloromethane, until the starting productdisappears. The solvent is evaporated off under reduced pressure and theproduct is purified by chromatography in order to produce 1.39 g of cisdiphenylmethyl3-hydroxy-1-[(2-propenyloxy)carbonyl]-4-piperidinepropanoate (M=423 g),i.e. an overall yield of 26%.

Then 1.2 g (2.83 mmoles) of the product obtained previously is dissolvedunder a nitrogen atmosphere in 23 ml of dichloromethane. Then 390 μl ofacetic acid then 860 μl of Bu₃SnH and 70 mg of Pd(PPh₃)₄ are added.

The solvent is evaporated off under reduced pressure in order to obtain3.82 g of crude product which is washed with petroleum ether. 1.27 g ofproduct is obtained which is filtered on silica with dichloromethane,then with a mixture of dichloromethane and methanol 95/5 then 90/10. Inthis way 0.87 g of cis diphenylmethyl 3-hydroxy-4-piperidinepropanoate(M=339 g) is obtained, i.e. a yield of 77%.

400 mg (1.00 mmole) of the compound obtained previously is dissolved in25 ml of dichloromethane, 80 μl of diphosgene (Cl₃COCOCl), 336 μl ofTEA, 144 mg of DMAP are added.

The reaction medium is left to react at ambient temperature for 5 hours30 minutes, then diluted in dichloromethane, followed by washing with a10% aqueous solution of tartaric acid, then with a buffer solution ofsodium phosphate at pH 7. The organic phase is dried over sodiumsulphate, then the solvent is evaporated off under reduced pressure. Inthis way 380 mg of crude product is obtained.

Purification is carried out by chromatography on silica, eluting with adichloromethane/ethyl acetate mixture 95/5 with 0.1% water.

184 mg of the title compound is obtained (M=365.43 g), i.e. a yield of50%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.60 to 1.88 (m): NCH ₂—CH ₂-CH; 2.48 (m): CH₂—CH ₂—CO; 2.78 (d)—2.90(m)—3.33 to 3.47 (m): CH ₂—N—CH ₂; 4.50 (d): CHO—CH₂; 6.89 (s):CO₂CH(C₆H₅)₂; 7.33 (m): (C₆H₅)₂.

IR (CHCl₃): 1784, 1734, 1600, 1585, 1496 cm⁻¹

MS (positive electrospray) m/z: [M]⁺=365

Example 1a cis-7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-4-propanoic acid

176 mg (0.482 mmoles) of the product obtained previously is dissolved in10 ml of acetone. 90 mg of Pd/C at 10% by weight is added.

The reaction medium is left to react under a hydrogen atmosphere atnormal pressure for 3 hours. Another 25 mg of catalyst is added and thereaction is left to continue for 1 hour 15 minutes.

The catalyst is filtered then the solvent is evaporated off underreduced pressure in order to obtain 146 mg of product.

The medium is reacted in 10 ml of acetone with 35 mg of Pd/C at 10% byweight under a hydrogen atmosphere and the reaction is left for 1 hourto complete.

The catalyst is then separated by filtration and the filtrate isevaporated under reduced pressure. 137 mg of crude product is obtainedwhich is crystallized from a mixture of ethyl ether and petroleum ether.In this way 75 mg of the sought product (M=199 g) is obtained, i.e. ayield of 78%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.30 to 1.63 (m) and 1.88 (m): NCH ₂—CH ₂—CH; 2.25 (t): CH₂—CH ₂—CO;3.06 (m) and 3.38 (m): CH ₂—N—CH ₂; 4.65 (d): C—CHO—CH₂; 12.08 (s):mobile H.

IR (Nujol): 1785, 1717 cm⁻¹

MS (FAB) m/z: [M+H]⁺=200; 159

Example 2 Trans diphenylmethyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetate

94 mg (0.259 mmoles) of the compound trans diphenylmethyl3-hydroxy-4-piperidine-acetate hydrochloride (M=361.87 g) (described inEur. J. Med. Chem—Chim. Ther—1982-17(6)531-5) and 7 ml ofdichloromethane are mixed under an inert atmosphere.

The reaction medium is cooled down using an ice bath and 19 μl ofdiphosgene is injected. Agitation is carried out for 25 minutes, then 72μl of TEA is injected. Agitation is carried out at ambient temperaturefor 30 minutes and the solvent is evaporated off under reduced pressure,followed by taking up in 7 ml of toluene.

36 μl of TEA then 31 mg of DMAP are added.

Heating is carried out for 15 minutes at 100° C., then the reactionmedium is left to return to ambient temperature, followed by washingtwice with 4 ml of 10% tartaric acid in water, then with 4 ml of watersaturated with sodium chloride.

After drying over magnesium sulphate, filtration is carried out and thesolvent is evaporated off under reduced pressure.

78 mg of an oil is obtained which is chromatographed on silica, elutingwith a 95/5 mixture of dichloromethane and ethyl acetate.

In this way 35.7 mg of expected compound (M=351.405 g) is obtained, inthe form of white crystals, i.e. a yield of 39%.

Example 2a trans-7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetic acid

38.7 mg (0.110 mmoles) of the product obtained in Example 2 as well as 2ml of acetone and 38 mg of Pd/C catalyst at 10% by weight are mixedtogether under an inert atmosphere.

The reaction medium is placed under a hydrogen atmosphere at normalpressure.

The reaction medium is left to react for 45 minutes, then the catalystis eliminated by filtration and the solvent is evaporated off underreduced pressure.

In this way 32.6 mg of crude product is obtained.

Recrystallization is carried out from ethyl ether in order to obtain14.2 mg of white crystals of expected compound (C₈H₁₀NO₄— M=185.181 g),i.e. a yield of 69%.

Example 3 cis diphenylmethyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetate

1.5 g (5.78 mmoles) oftrans-1-[(1,1-dimethylethoxy)carbonyl]-3-hydroxy-4-piperidineacetic acid(described in Eur. J. Med. Chem—Chim. Ther—1982-17(6)531-5), 7 ml ofdichloromethane, 3.03 g of triphenylphosphine and 22 ml oftetrahydrofuran are mixed together.

A solution of 0.91 ml of DEAD in 2.5 ml of tetrahydrofuran is added. Thereaction medium is left to react for 3 hours 20 minutes, then 8.7 ml of1N soda is added and agitation is carried out for 1 hour 15 minutes.

The reaction mixture is extracted twice with ethyl acetate, then the pHis adjusted to 2 with 2N hydrochloric acid, followed by extracting threetimes with ethyl acetate.

The organic phases are combined and washed with water saturated insodium chloride, then dried over magnesium sulphate, filtered and thesolvent is evaporated off under reduced pressure.

In this way 1.37 g of white crystals of 1,1-dimethylethyl(3a.alpha.,7a.alpha.)-hexahydro-2-oxo-furo[2,3-c]pyridine-6(2H)-carboxylate(C₁₂H₂₁NO₅-M=259.304 g) is obtained, i.e. a yield of 91%.

1.37 g (5.28 mmoles) of the compound obtained previously and 32 ml ofdichloromethane are mixed together under an inert atmosphere.

An excess of a solution of diphenyldiazomethane in dichloromethane isintroduced, until the starting product disappears.

Then the solvent is evaporated off under reduced pressure and in thisway 2.81 g of crude product is obtained which is purified bychromatography on silica, using dichloromethane, then a 95/5 mixture ofdichloromethane/ethyl acetate as eluent.

2.00 g of white crystals of cis diphenylmethyl1-[(1,1-dimethylethoxy)carbonyl]-3-hydroxy-4-piperidineacetate isobtained, (M=425.528 g), i.e. a yield of 89%.

0.6 g (1.41 mmoles) of the compound obtained previously and 1.93 ml of asolution of hydrogen chloride in methanol at 7.3 mol/l is introduced,

Agitation is carried out at ambient temperature and after 15 minutes, 1ml of dichloromethane is added.

After another 15 minutes, the reaction medium is evaporated underreduced pressure.

Dichloromethane is again added then, evaporation is carried out. Thisoperation is repeated several times.

Then the product is crystallized from ethyl ether.

In this way 0.44 g of the hydrochloride of cis diphenylmethyl3-hydroxy-4-piperidineacetate is obtained of molecular formula,C₂₀H₂₃NO₃, HCl (M=361.871 g), i.e. a yield of 86%.

This reaction also leads to the formation of variable quantities of thelactone hydrochloride of(3a.alpha.,7a.alpha.)-hexahydro-furo[2,3-c]pyridin-2(3H)-one, (M=177.6g).

0.28 g (0.77 mmoles) of compound C₂₀H₂₃NO₃, HCl obtained previously and19 ml of dichloromethane are mixed together under an inert atmosphere.

60 μl of diphosgene is added at 0° C. and agitation is carried out.After 25 minutes 0.32 ml of TEA is introduced. Then 94 mg of DMAP isadded and the reaction medium is left to return to ambient temperature.

Agitation is carried out for 4 hours 15 minutes, followed by washingsuccessively with a 10% aqueous solution of tartaric acid then withwater saturated with sodium chloride, followed by drying over magnesiumsulphate, filtering and the solvent is evaporated off under reducedpressure.

In this way 0.265 g of expected compound of molecular formula C₂₁H₂₁NO₄(M=351.405 g) is obtained, i.e. a yield of 98%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.82 (m): NCH₂—CH ₂; 2.30 to 2.70 (m): CO—CH₂—CH; 2.93 (d)—2.99 (dt) and3.45 (m): CH ₂—N—CH ₂; 4.60 (d): CH—CHO—CH₂; 6.87 (s): CO₂CH(C₆H₅)₂;7.10 to 7.35 (m): (C₆ H ₅)₂. IR (CHCl₃)=1786, 1734; 1600, 1587, 1496cm⁻¹.

MS (SIMS) m/z: [M+Na]⁺=374⁺.

Example 3a cis-7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetic acid

55 mg (0.156 mmoles) of the product obtained in Example 3, 3 ml of ethylacetate and 55 mg of Pd/C catalyst at 10% by weight are mixed together.

The reaction medium is placed under a hydrogen atmosphere at normalpressure.

The reaction medium is left to react for 1 hour 30 minutes, then thecatalyst is filtered out and the solvent is evaporated off under reducedpressure.

In this way 38 mg of crude product is obtained which is crystallizedfrom a mixture of pentane and ethyl ether.

In this way 16 mg of white crystals of expected compound (M=185.181 g)is collected, i.e. a yield of 55%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.63 to 1.86 (m) and 1.91 (m): NCH₂—CH ₂; 2.27 to 2.49 (m) and 2.54(dd): CO—CH ₂—CH; 2.98 (d) and 3.54 (d): CH ₂—N—CH₂—CH₂; 3.04 (dt) and3.41 (dd): CH₂—N—CH ₂—CH₂; 4.71 (d):

CH—CHO—CH₂

IR (Nujol): 1784, 1734, 1686 cm⁻¹.

MS (SIMS) m/z: [M+H]⁺=186⁺, 167⁺.

Example 3b cis methyl 7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetate

78 mg (0.421 mmoles) of the compound obtained in Example 3a is thendissolved in 1 ml of dichloromethane.

An excess of diazomethane is added dropwise until a yellow colourationsubsists, then the solvent is evaporated off under reduced pressure.

In this way 80 mg of crude product is obtained which is purified bychromatography on silica, eluting with a dichloromethane/ethyl acetatemixture 95/5.

In this way 8.2 mg of expected compound (M=199.208 g) is obtained i.e. ayield of 10%.

Example 4 cis-7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetonitrile

67 mg (0.38 mmoles) of the hydrochloride of(3a.alpha.,7a.alpha.)-hexahydro-furo[2,3-c]pyridin-2(3H)-one, (M=177.6g) prepared in Example 3 is dissolved in 1 ml of a solution of ammoniaat 4.17 mol/l in methanol.

Agitation is carried out for 5 hours, the solvent is evaporated offunder reduced pressure, then another 1 ml of the solution of ammonia inmethanol is added and the reaction is left to continue for 18 hours.

The solvent is evaporated off under reduced pressure and in this way 79mg of cis-3-hydroxy-4-piperidineacetamide of molecular formula C7H14O2N2(M=158 g) is obtained.

75 mg of the compound obtained above is mixed under an inert atmospherein solution in 9 ml of dichloromethane.

The reaction medium is cooled down with an ice bath and 30 μl ofdiphosgene is introduced.

The reaction medium is maintained at 0-5° C. for 40 minutes, then 0.16ml of TEA is introduced and after 5 minutes, 46 mg of DMAP isintroduced.

Agitation is carried out for 4 hours at ambient temperature, followed bywashing twice with 2 ml of 10% tartaric acid in water, then with 2 ml ofa saturated aqueous solution of sodium chloride.

After drying over MgSO4 and filtering, the solvent is evaporated offunder reduced pressure. In this way 35 mg of crude product is obtainedwhich is taken up in an ethyl acetate and dichloromethane mixture 30/70.The impurities are filtered out and the filtrate is evaporated underreduced pressure.

In this way 23 mg of expected compound (M=166.18 g) is obtained in theform of an oil, i.e. a yield of approximately 26%.

IR (Nujol): 2241, 1777 cm⁻¹.

MS (EI) m/z: [M]⁺=166, 137, 82, 55, 42.

Example 5 3-benzoyl-1,3-diazabicyclo[2.2.1]heptan-2-one

1-01 g (5.43 mmoles) of 1,1-dimethylethyl3-amino-1-pyrrolidinecarboxylate (M=186.25 g) (described in the PatentApplication WO 9801426) and 10 ml of dichloromethane are mixed under aninert atmosphere, the solution is cooled down to 0° C., then 0.76 ml ofTEA. is added dropwise.

Agitation is carried out for 15 minutes whilst maintaining thetemperature at 0° C., then 0.63 ml of benzoyl chloride is added.

The reaction medium is left to return to ambient temperature, thendiluted by adding 10 ml of dichloromethane, followed by washing with a10% aqueous solution of tartaric acid, then with 10 ml of water, dryingover magnesium sulphate, filtering and the dichloromethane is eliminatedby evaporation under reduced pressure.

In this way 1.30 g of 1,1-dimethylethyl3-(benzoylamino)-1-pyrrolidinecarboxylate (M=292.36 g) is obtained inthe form of a yellow oil. The corresponding yield is 82%.

1.30 g (4.46 mmoles) of this compound is mixed with 10 ml of methanol.

The solution is cooled down to 0° C., then 6.12 ml of a solution ofhydrogen chloride at 7.3 moles/l in methanol is introducedprogressively.

Then the solvent is evaporated off under reduced pressure.

In this way 1.01 g of N-(3-pyrrolidinyl)-benzamide hydrochloride(M=226.707 g) is obtained in the form of oil brown, i.e. a yield closeto 100%.

1.01 g (4.46 mmoles) of the compound obtained previously, as well as 10ml of dichloromethane are mixed under an inert atmosphere.

The reaction medium is cooled down to 0° C., then 1.36 ml of TEA isadded dropwise.

Agitation is carried out for 15 minutes, then 1.44 ml of diphosgene isadded dropwise.

The reaction medium is maintained at 0° C. for 30 minutes, then left toreturn to ambient temperature, followed by diluting withdichloromethane, washing with a 10% aqueous solution of tartaric acid,then with water, drying over magnesium sulphate, filtering andconcentrating the solvent by evaporating under reduced pressure in orderto obtain 0.615 g of crude product.

Purification is carried out by chromatography on silica eluting with adichloromethane/acetone mixture 90/10.

In this way 0.320 g of the chloride of3-(benzoylamino)-1-pyrrolidinecarboxylic acid is recovered whichcrystallizes. The corresponding yield is 28%.

Then 0.585 g (2.31 mmoles) of the preceding compound is dissolved underan inert atmosphere in 18 ml of tetrahydrofuran.

The solution is cooled down to −78° C., then 2.55 ml of a 1 M solutionof lithium bis(trimethyl-silyl)amide in tetrahydrofuran is addeddropwise.

A yellow solution is obtained which is maintained at −78° C. for 20minutes, then agitation is continued for 1 hour whilst allowing thetemperature to rise. 350 μl of acetic acid, then 5 ml of a 10% solutionof tartaric acid in water are added at 0° C., followed by diluting withethyl acetate then washing with a 10% solution of tartaric acid thenwith a solution of phosphate buffer at pH=7, then with water.

The organic phase is dried over magnesium sulphate, followed byfiltering and concentrating the solvent by evaporating under reducedpressure.

In this way 0.315 g of crude product is obtained in the form of a yellowsolid.

This crude product is purified by chromatography on silica eluting witha dichloromethane and ethyl acetate mixture 90/10.

In this way 0.140 g of expected compound C₁₂H₁₂N₂O₂, (M=216.24 g) isrecovered in the form of a white solid, i.e. a yield of 28%.

IR (CHCl₃): 1801, 1775, 1675; 1620, 1603, 1582 cm⁻¹.

MS (positive electrospray) m/z: [M]⁺=216, 105, 77.

Example 6 Potassium salt oftrans-6-[(phenylmethoxy)carbonyl]-2-oxo-1,3-diazabicyclo[2.2.1]heptan-3-aceticacid

1 g (3.12 mmoles−M=186.25 g) of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-amino-1,2-pyrrolidinedicarboxylate (described in J.Org. Chem. 1991, 56, 3009-3016), 10 ml of tetrahydrofuran, 560 μl ofallyl bromoacetate and 660 μl of TEA are mixed together.

The reaction medium is left to react under agitation at ambienttemperature for 14 hours, then for 3 hours at 50° C., followed bydiluting with ethyl acetate and washing with a 10% aqueous solution oftartaric acid, then with a saturated aqueous solution of sodiumchloride.

The organic phase is dried over magnesium sulphate, filtered then thesolvent is evaporated off under reduced pressure.

In this way 1.21 g of crude product is obtained which is purified bychromatography on silica, eluting with a 80/20 mixture ofdichloromethane and ethyl acetate.

0.99 mg of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-[[[(2-propenyloxy)carbonyl]methyl]amino]-1,2-pyrrolidinedicarboxylate of molecular formula C₁₂H₃₀N₂O₆ (M=418 g) is obtained. 6ml of a 4 M solution of hydrogen chloride in ethyl acetate is addedunder a nitrogen atmosphere and at 0° C. to 0.99 g (2.36 mmoles) of thecompound obtained previously. The reaction medium is left to react atambient temperature for 15 minutes.

The solvent is evaporated off under reduced pressure. A crude product isobtained which is crystallized from ethyl ether in order to obtain 0.95g of the dihydrochloride of trans phenylmethyl4-[[[(2-propenyloxy)carbonyl]methyl]amino]-2-pyrrolidinecarboxylate, ofmolecular formula C₁₇H₂₃N₂O₄, 2HCl (M=394 g).

0.5 g of this product is dissolved in 20 ml of dichloromethane and 1.3ml of 2N soda and 3 ml of water are added. The reaction medium is leftto settle, followed by extracting with dichloromethane, drying overmagnesium sulphate, then filtering and the solvent is evaporated offunder reduced pressure.

In this way 339 mg of free diamine is obtained. The corresponding yieldis 83%.

100 mg (0.314 mmoles) of the diamine obtained previously is dissolved in5 ml of acetonitrile at 0° C. and under a nitrogen atmosphere.

21 μl of diphosgene is added. After 15 minutes of contact, this solutionis added, under a nitrogen atmosphere and over 4 hours, to a mixturecontaining 38 mg of DMAP, 88 μl of TEA in 10 ml of acetonitrile heatedto 70° C.

After the addition has ended, the reaction mixture is heated again forone hour, then cooled down, diluted with ethyl acetate and washedsuccessively with a 10% aqueous solution of tartaric acid, then with asaturated aqueous solution of sodium chloride. After drying over sodiumsulphate, filtering and evaporating the solvents under reduced pressure,58 mg of crude product is obtained. This product is purified bychromatography on silica eluting with a dichloramethane/ethyl acetatemixture 8/2 in order to produce 19 mg of trans 2-propenyl6-[(phenylmethoxy)carbonyl]-2-oxo-1,3-diazabicyclo[2.2.1]heptan-3-acetateof molecular formula C₁₈H₂₀N₂O₅ (M=344.57 g), i.e. a yield of 17%.

Then 24 mg (0.069 mmoles) of the preceding compound is dissolved in 250μl of dichloromethane. 3 mg of Pd(PPh₃)₄ is introduced under a nitrogenatmosphere, then 150 μl of a 0.5 M solution of potassiumethyl-2-hexanoate in ethyl acetate is added. After a few minutes, aprecipitate forms which is centrifuged and washed twice with 500 μl ofethyl acetate.

24 mg of expected compound C₁₅H₁₅KN₂O₅ (M=342 g) is obtained, i.e. aquantitative yield.

1H NMR

In DMSO, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.83 (ddd) and 2.56: N—CH₂—CHN—CH ₂; 2.50 and 2.79 (d): N—CH ₂—CHN—CH₂;3.23 (d) and 3.41 (d): ═C—N—CH ₂—C═O; 3.62 (ddd): O═C—CHN—CH ₂; 4-0.13(s): N—CH₂—CHN—CH₂; 5.16 (s): ═C—O—CH ₂—C₆H₅; 7.38 (m): C₆H₅—CH₂.

MS (positive electrospray) m/z: [2MK+H]⁺=723, [2MK+Na]⁺=707,[MK+K]⁺=381, [MK+Na]⁺=365; [MK+H]⁺=343.

Example 7 trans methyl3-benzoyl-2-oxo-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate

0.471 g (1.93 mmole) of trans 1-(1,1-dimethylethyl) and 2-methyl4-amino-1,2-pyrrolidinedicarboxylate (described in J. Org. Chem. 1991,56, 3009-3016) and 3.5 ml of dry dichloromethane to dissolve it aremixed under a nitrogen atmosphere.

The solution is cooled down to 0° C., then 269 μl of TEA is addeddropwise.

Agitation is carried out for 15 minutes whilst maintaining at 0° C.,then 224 μl of benzoyl chloride is added dropwise.

The reaction medium is left and the temperature returns to 20° C. overone hour, followed by diluting with 30 ml of dichloromethane, washingwith a 10% aqueous solution of tartaric acid, then with a saturatedsolution of sodium bicarbonate, then with water, drying over magnesiumsulphate, filtering, concentrating by evaporating the dichloromethaneunder reduced pressure.

In this way 0.6 g of a yellow oil is obtained which is purified bychromatography on silica using a dichloromethane/methanol mixture 99/1as eluent.

In this way 0.499 g of trans 1-(1,1-dimethylethyl) and 2-methyl4-(benzoylamino)-1,2-pyrrolidine dicarboxylate of molecular formulaC₁₈H₂₄N₂O₅ (M=348 g) is recovered, i.e. a yield of 74%.

0.400 g (1.15 mmole) of the compound obtained previously with 3 ml ofethyl acetate to dissolve the compound are mixed under a nitrogenatmosphere, then the solution is cooled down to 0° C., 2.89 ml of asolution of 4 mole/l of HCl in ethyl acetate is added.

At the end of 15 minutes, agitation is continued at ambient temperaturefor 1 hour.

Then the solvent is eliminated by evaporation under reduced pressure.

In this way 0.350 g of the hydrochloride of trans methyl4-(benzoylamino)-2-pyrrolidinecarboxylate of molecular formulaC₁₃H₁₅N₂O₃, HCl (M=284.744 g) is obtained in the form of a beige solid.

0.327 g (1.15 mmole) of the compound obtained previously, placed under anitrogen atmosphere, is mixed with 4 ml of dichloromethane.

The suspension is cooled down to 0° C., then 352 μl of TEA is added.Agitation is carried out for 15 minutes at 0° C., then 138 μl ofdiphosgene is added. Agitation is continued for 5 minutes at 0° C., thenthe reaction mixture is left to return to ambient temperature and leftto react for 30 minutes, followed by diluting with dichloromethane andwashing with a 10% aqueous solution of tartaric acid, then with waterand drying over magnesium sulphate.

After filtering, the solvent is eliminated by evaporation under reducedpressure. In this way 0.360 g of crude product is obtained which ispurified by chromatography on silica eluting with adichloromethane/acetone mixture 95/5.

In this way 93.7 mg of the hydrochloride of trans methyl4-(benzoylamino)-1-(chlorocarbonyl)-2-pyrrolidine carboxylate(C₁₄H₁₄N₂O₄, HCl (M=310.74 g) is recovered, i.e. a yield of 26%.

93.7 mg (0.301 mmole) of the compound obtained previously, is mixedunder a nitrogen atmosphere, with 3 ml of tetrahydrofuran. Thetemperature of the solution is lowered to −78° C., then 332 μl oflithium bis(trimethylsilyl)amide in a 1M solution in tetrahydrofuran isadded dropwise and the reaction medium is maintained at −78° C. foranother 5 minutes.

Agitation is carried out for 30 minutes at ambient temperature.

Then the solution is cooled down to 0° C., and 55 μl of acetic acid isadded. 20 ml of ethyl acetate and 3 ml of a phosphate buffer at pH=7.0are added. The reaction medium is left to settle, followed by washingwith water, drying over magnesium sulphate, filtering and concentratingby evaporation. In this way 76 mg of a foam is obtained which ispurified by chromatography on silica eluting with adichloromethane/acetone mixture 97/3.

5 mg of pure expected compound, of molecular formula (C₁₄H₁₄N₂O₄, HCl(M=274.279 g), is recovered i.e. a yield of 6%.

IR (CHCl₃): 1805, 1779, 1743, 1669; 1603, 1589, 1486 cm⁻¹

MS (EI) m/z: [M]⁺=274, 215, 169, 105, 77.

Example 7a trans phenylmethyl3-benzoyl-2-oxo-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate

The operation is carried out in a similar fashion to that indicated inExample 7, starting from 0.92 g of trans 1-(1,1-dimethylethyl) and2-phenylmethyl 4-amino-1,2-pyrrolidinedicarboxylate (described in J.Org. Chem. 1991, 56, 3009-3016) in order to obtain the expected compoundwith an overall yield of 5.4% over 4 stages.

Example 8 trans phenylmethyl2-oxo-3-(phenylsulphonyl)-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate

2.97 g (9.26 mmoles) of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-amino-1,2-pyrrolidinedicarboxylate (described in J.Org. Chem. 1991, 56, 3009-3016) of molecular formula C₁₇H₂₄N₂O₄(M=320.392 g) is mixed under a nitrogen atmosphere and 25 ml ofdichloromethane is added. The reaction medium is cooled down to 5° C.and 1.3 ml of TEA is added. Agitation is carried out for 10 minutes andthen 1.63 g of benzenesulphonyl chloride is added.

The reaction medium is left under agitation at 5° C. for 15 minutes,then the temperature of the reaction medium is allowed to rise to 20° C.for a duration of 45 minutes, followed by diluting with dichloromethane,washing with a 10% aqueous solution of tartaric acid, then withphosphate buffer at pH=7.0, then with a saturated aqueous solution ofsodium chloride, drying over magnesium sulphate and the solvent isevaporated off under reduced pressure.

In this way 4.5 g of crude product is obtained which is chromatographedon silica eluting with a 90/10 mixture of dichloromethane and ethylacetate.

In this way 4.06 g of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-[(phenylsulphonyl)amino]-1,2-pyrrolidinedicarboxylateof molecular formula C₂₃H₂₈N₂O₆S (M=460, 552 g) is recovered, whichcorresponds to a yield of 95%.

3.83 g (8.31 mmoles) of the sulphonamide obtained previously is mixedwith 10 ml of anhydrous methanol.

The solution is cooled down to 0° C. and 8.2 ml of a solution of 10mol/l of hydrochloric acid in methanol is added at this temperature.

Agitation is maintained at 0° C. for 5 minutes, then the temperature isallowed to rise until at ambient temperature.

After 30 minutes, the methanol is evaporated off under reduced pressure,followed by taking up several times in methanol then in dichloromethane.Then the hydrochloride is crystallized from ethyl ether.

In this way 3.2 g of the hydrochloride of trans phenylmethyl4-[phenylsulphonyl)amino]-2-pyrrolidinecarboxylate, of molecular formulaC₁₈H₂₀N₂O₄S, HCl (M=396.896 g) is obtained, which corresponds to a yieldof 96%.

2.78 g (7 mmoles) of the hydrochloride obtained previously under aninert atmosphere is mixed with 28 ml of dichloromethane.

The reaction medium is cooled down to about 0-5° C., then 2.15 ml of TEAis added.

Agitation is continued for 15 minutes at a temperature comprised between0 and 5° C., then 0.46 ml of diphosgene is added.

The reaction medium is maintained at this temperature for 4 minutes,then a 10% aqueous solution of tartaric acid is added, followed bydiluting with dichloromethane, decanting, washing with a saturatedaqueous solution of sodium chloride, drying over magnesium sulphate, andconcentrating under reduced pressure.

In this way 3.1 g of a yellow oil is obtained which is purified bychromatography on silica, eluting with a dichloromethane/ethyl acetatemixture 9/1.

1.82 g of trans phenylmethyl1-(chlorocarbonyl)-4-[(phenylsulphonyl)amino]-2-pyrrolidinecarboxylateof molecular formula C₁₉H₁₉ClN₂O₅S (M=422.89 g) is recovered, whichcorresponds to a yield of 61%.

1.81 g (4.28 mmoles) of the carbamoyl chloride obtained previously ismixed under an inert atmosphere with 31 ml of tetrahydrofuran.

The solution obtained is cooled down to −70° C., then 4.7 ml of a 1Msolution of lithium bis(trimethylsilyl)amide in tetrahydrofuran is addedat this temperature over 10 minutes.

Agitation is carried out for 45 minutes at −70° C., then the temperatureof the reaction medium is allowed to rise to about 0° C. The reactionmedium is maintained at this temperature for 2 hours 30 minutes.

Then 295 μl of acetic acid is added, followed by diluting withdichloromethane, washing with a 10% aqueous solution of tartaric acid,with a solution of phosphate buffer at pH=7 and with a saturated aqueoussolution of sodium chloride, drying over magnesium sulphate andconcentrating to dryness under reduced pressure.

The crude product is purified by chromatography on silica, eluting witha dichloromethane/ethyl acetate mixture 95/5.

In this way 244 mg of expected compound of molecular formula C₁₉H₁₈N₂O₅S(M=386.429 g) is obtained, which corresponds to a yield of 14%.

1H NMR

In CDCl3, at 400 MHz, chemical shifts of the peaks in ppm andmultiplicity:

2.15 (m): O═C—CH—CH ₂; 2.85 (d) and 3.08 (d): O═C—N—CH ₂; 3.62 (m):O═C—CH—N—CH₂; 4.94 (s): O₂S—N—CH—CH₂; 5.16: CO₂CH ₂C₆H₅; 7.34 (m): C₆ H₅; 7.57 (m)-7.68 (m) and 8.03 (m): SO₂C₆ H ₅.

IR (CHCl₃): 1780, 1743; 1586, 1499 cm⁻¹

MS (positive electrospray) m/z: [2M+Na]⁺=795;

[M+Na+CH₃CN]⁺=450; [M+Na]⁺=409; [M+H]⁺=387.

Example 9 trans phenylmethyl3-benzoyl-4-methyl-2-oxo-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate

18.69 g (58.52 mmoles) of 1-(1,1-dimethyl-ethyl) and2-(phenylmethyl)4-oxo-1,2 pyrrolidinedicarboxylate (described in Chem.Pharm. Bull. 43(8)1302-1306 (1995)) of molecular formula C₁₇H₂₁NO₅(M=319.361 g) and 500 ml of anhydrous ethyl ether are mixed togetherunder an inert atmosphere.

A suspension of 10 g of CeCl3 in 50 ml of anhydrous ethyl ether is addedto the solution obtained.

The suspension is agitated for 30 minutes to 20° C., then is cooled downto −60° C.

Then 20 ml of 3 M solution of MeMgBr in ethyl ether is added.

The reaction medium is left to react for 1 hour at −60° C., then thetemperature is allowed to rise to 0° C. over 30 minutes, followed byneutralizing with a 10% aqueous solution of NH₄Cl, extracting withdichloromethane, filtering, washing the organic phase with water, dryingover magnesium sulphate, and concentrating to dryness under reducedpressure.

In this way 19.33 g of an oil is obtained which is purified bychromatography on silica eluting with adichloromethane/tbutylmethyl-ether mixture 90/10.

7.21 g of cis 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-hydroxy-4-methyl-1,2-pyrrolidine dicarboxylate ofmolecular formula C₁₈H₂₅NO₅ (M=335.404 g) is obtained, i.e. a yield of36%, as well as 2.5 g of the alcohol epimer.

3.17 g (9.45 mmoles) of the compound obtained previously and 70 ml ofdichloromethane are mixed together under an inert atmosphere. Thereaction medium is cooled down to 5° C. and 2.3 ml of TEA, then 1.28 mlof methane sulphonyl chloride are added dropwise.

Agitation is carried out for 45 minutes at 5° C., followed by washingwith a 10% aqueous solution of tartaric acid, then with a solution ofphosphate buffer at pH 7, then with water.

The organic phase is dried over magnesium sulphate and concentrated todryness under reduced pressure.

In this way 3.9 g of an oil is obtained which is purified bychromatography on silica eluting with a dichloromethane/ethyl acetatemixture 90/10.

2.75 g of cis 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-methyl-4-[(methylsulphonyl)oxy]-1,2-pyrrolidinedicarboxylate of molecular formula C₁₉H₂₇NO₇S (M=413.494 g) is recoveredwhich corresponds to a yield of 70%.

A solution of 2.54 g (6.14 mmoles) of the mesylate obtained previouslyin 40 ml of dimethylformamide is prepared.

Then, 519 mg (7.98 mmoles) of NaN₃ is added at 20° C. followed byheating at 50° C. for 2 hours. After cooling down, the reaction mediumis poured into 250 ml of water and extracted with 250 ml ofdichloromethane. The organic phase is washed with water, dried overmagnesium sulphate and evaporated to dryness under reduced pressure.

2.4 g of crude product is obtained which is purified by chromatographyon silica, eluting with a dichloromethane/ethyl acetate mixture 95/5.

In this way 1.66 g of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-azido-4-methyl-1,2-pyrrolidine dicarboxylate ofmolecular formula C₁₈H₂₄N₄O₄ (M=360.42 g) is recovered (titringapproximately 30% by weight), which corresponds to a yield ofapproximately 25%.

1.85 g of the azide obtained previously (i.e. approximately 1.7 mmole)is dissolved in 18 ml of toluene.

Then, 1.38 ml of Bu3SnH and 84 mg of AIBN are added at 20° C.

The reaction medium is taken to 75° C. and maintained at thistemperature for 2 hours.

The toluene is evaporated off and redissolving is carried out in ethylacetate. A saturated aqueous solution of potassium fluoride is added andagitation is carried out for 30 minutes at ambient temperature, followedby filtering on clarcel, leaving to settle and drying the organic phaseover magnesium sulphate.

After evaporation of the solvent under reduced pressure, 3 g of an oilis obtained which is chromatographed on silica, eluting with adichloromethane/methanol mixture 9/1.

560 mg of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)-4-amino-4-methyl-1,2-pyrrolidine dicarboxylate ofmolecular formula C₁₈H₂₆N₂O₄ (M=334.419 g) is recovered. The yield istherefore quantitative.

578 mg (1.72 mmoles) of the amine obtained previously is mixed under aninert atmosphere in 30 ml of dichloromethane.

The reaction medium is cooled down to 5° C. and 290 μl of TEA, then 240μl of benzoyl chloride are added dropwise.

Agitation is continued at 5° C. for 30 minutes, followed by dilutingwith dichloromethane, washing with a 10% aqueous solution of tartaricacid, with a saturated aqueous solution of sodium carbonate, then withwater, drying the organic phase over magnesium sulphate, and evaporatingthe solvent under reduced pressure.

In this way 950 mg of an oil is obtained which is purified bychromatography eluting with a dichloromethane/ethyl acetate mixture90/10.

In this way 732 mg of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-(benzoylamino)-4-methyl-1,2-pyrrolidinedicarboxylateof molecular formula C₂₅H₃₀N₂O₅. (M=438.528 g) is recovered, whichcorresponds to a yield of 97%.

636 mg (1.45 mmoles) of the amide obtained previously is dissolved in1.9 ml of ethyl acetate, the reaction medium is cooled down to about0-5° C. with an ice bath, then 3.2 ml of a solution of hydrogen chlorideat 4.6 mol/l in ethyl acetate is added.

The temperature of the reaction medium is left to rise to 20° C., thenafter 1 hour, the solvent is evaporated off under reduced pressure.

Then the hydrochloride crystallizes from ethyl ether.

In this way 570 mg of the hydrochloride of trans phenylmethyl4-(benzoylamino)-4-methyl-2-pyrrolidine carboxylate of molecular formulaC₂₀H₂₂N₂O₃, HCl (M=374.87 g) is recovered, in the form of a whitepowder. The yield is therefore quantitive.

100 mg (0.267 mmole) of the hydrochloride obtained previously isdissolved under an inert atmosphere in 1.5 ml of dichloromethane.

The reaction medium is cooled down to about 0-5° C., then 90 μl of TEAis added.

Agitation is carried out for 15 minutes at 5° C., then 20 μl ofdiphosgene is added.

Agitation is continued for 30 minutes at 5° C.

Then, the reaction medium is treated with a 10% aqueous solution oftartaric acid, followed by extracting with dichloromethane, washing theorganic phase with a saturated aqueous solution of sodium chloride,drying over magnesium sulphate, and evaporating the solvent underreduced pressure.

In this way 130 mg of an oil is obtained which is purified bychromatography on silica eluting with a dichloromethane/ethyl acetatemixture 9/1.

Then 72 mg of trans phenylmethyl4-(benzoylamino)-1-(chlorocarbonyl)-4-methyl-2-pyrrolidine carboxylateof molecular formula C₂₁H₂₁N₂O₄Cl (M=400.865 g) is recovered, whichcorresponds to a yield of 67%.

373 mg (0.930 mmole) of the compound obtained previously is dissolved in9 ml of tetrahydrofuran.

The solution is then cooled down to −70° C. and 1 ml of a 1 M solutionof lithium bis(trimethylsilyl)amide in the tetrahydrofuran is added over5 minutes.

The reaction medium is left to heat up to 0° C. over 45 minutes, then 69μl of acetic acid is added, followed by diluting with dichloromethane,washing with a 10% aqueous solution of tartaric acid, then with asolution of phosphate buffer at pH=7.0 and with a saturated aqueoussolution of sodium chloride.

The organic phase is dried over magnesium sulphate, followed byconcentrating to dryness under reduced pressure, in order to obtain 330mg of a crude product which is purified by chromatography on silica,eluting with a dichloromethane/ethyl acetate mixture 98/2 containing0.1% by volume of TEA.

In this way 123 mg of expected compound of molecular formula C₂₁H₂₀N₂O₄(M=364.404 g) is recovered, which corresponds to a yield of 36%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.76 (s): CH3; 2.11 (dd) and 2.73 (ddd): N—CH—CH ₂; 2.93 (dt) and 3.00(d): N—CH ₂; 3.96 (ddd): N—CH—CH₂; 5.21: CO₂CH ₂C₆H₅; 7.36 (m): CH₂C₆ H₅; 7.43 (t) and 7.57 (tt) and 7.72 (d): COC₆ H ₅.

IR (CHCl₃): 1776, 1745, 1682; 1601, 1580, 1498 cm⁻¹

MS (positive electrospray) m/z: [2M+Na]⁺=751; [2M+H]⁺=729; [M+Na]⁺=387;[M+H]⁺=365

Example 10 1-propenyltriphenylphosphonium salt of trans phenylmethyl2-oxo-3-(sulphooxy)-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate

15 g (46.71 mmoles) of cis 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-hydroxy-1,2-pyrrolidinedicarboxylate (commercialproduct) of molecular formula C₁₇H₂₃NO₅ (M=321.377 g) is dissolved underan inert atmosphere in 225 ml of anhydrous dichloromethane.

5.42 ml of 2,6-lutidine is added to the solution. The reaction medium iscooled down to −70° C., then, 8.25 ml of trifluoromethanesulphonicanhydride is introduced over 5 minutes.

Agitation is carried out for 10 minutes at −70° C. then 4.43 g ofO-allyl-hydroxyl-amine is introduced at −70° C.

Then the reaction mixture is left at ambient temperature for 27 hours,followed by diluting with dichloromethane, then washing with a 10%aqueous solution of tartaric acid, with a saturated aqueous solution ofNaHCO₃, and with water.

The organic phase is dried over sodium sulphate, and the solvent isevaporated off under reduced pressure.

In this way 23 g of a crude oil is obtained which is purified bychromatography on silica, the eluent being successively adichloromethane/ethyl acetate mixture 95/5, 90/10, then 80/20.

7.18 g of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-[(2-propenyloxy)amino]-1,2-pyrrolidine dicarboxylateof molecular formula C₂₀H₂₈N₂O₅ (M=376.456 g) is recovered, whichcorresponds to a yield of 40%.

3.25 g (8.63 mmoles) of the compound obtained previously is dissolved in3.5 ml of ethyl acetate.

The reaction medium is cooled down to about 0-5° C., then 19 ml of asolution of 4.6 mol/l of hydrogen chloride in ethyl acetate is added.

The reaction medium is left to react whilst agitating at about 0-5° C.for 40 minutes.

The solvent is evaporated off under reduced pressure, followed by takingup several times in diethyl ether, whilst drawing off the liquidsupernatant.

In this way 2.54 g of a hydrochloride is obtained in the form of a whiteprecipitate, which is dissolved in 55 ml of dichloromethane underagitation. 7.3 ml of 2N soda is added. After decanting, the organicphase is dried over sodium sulphate.

The dichloromethane is evaporated off under reduced pressure.

In this way 2.12 g of trans phenylmethyl4-[(2-propenyloxy)amino]-2-pyrrolidinecarboxylate of molecular formulaC₁₅H₂₀N₂O₃ (M=276.337 g) is obtained in the form of an oil i.e. a yieldof 89%.

4.14 g (15 mmoles) of the compound obtained previously is dissolvedunder an inert atmosphere in 1.5 l of acetonitrile.

The reaction medium is cooled down to about 0-5° C. and 1.14 ml ofdiphosgene is added. Agitation is carried out for 15 minutes whilstmaintaining at 0-5° C., then 4.6 ml of TEA, and 1.83 g of DMAP in 80 mlof acetonitrile are added successively.

The temperature is allowed to rise to ambient temperature and thereaction medium is left to react for 26 hours, then half the solvent isevaporated off under reduced pressure, followed by treating with a 10%aqueous solution of tartaric acid, then extracting with dichloromethane.The organic phase is washed with a saturated aqueous solution of sodiumchloride, dried over magnesium sulphate and the solvent is evaporatedoff under reduced pressure.

In this way 43 g of crude product is obtained which is purified bychromatography on silica eluting with a dichloromethane/ethyl acetatemixture 90/10 containing 0.1% of TEA.

312 mg of trans phenylmethyl2-oxo-3-(2-propenyloxy)-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate ofmolecular formula C₁₆H₁₈N₂O₄ (M=302.33 g) is recovered which correspondsto a yield of 7%.

70.2 mg (0.232 mmole) of the compound obtained previously is dissolvedunder an inert atmosphere in 2.3 ml of dichloromethane. Then 26.5 μl ofacetic acid and 134 mg of Pd(P(Ph)₃)₄ are introduced.

The reaction medium is left to react for 40 minutes at ambienttemperature, then the temperature is lowered to −20° C. and 2.96 ml of asolution of SO3-pyridine complex at 0.314 mol/l is added. The reactionmedium is left to react for 2 hours and 30 minutes then dichloromethaneis added followed by evaporating under reduced pressure, taking up in 40ml of dichloromethane and washing with 5 ml of water. The organic phaseis separated and dried over sodium sulphate, then the solvent isevaporated off under reduced pressure.

In this way 280 mg of crude product is obtained which is purified bychromatography on silica, eluting successively with adichloromethane/acetone mixture 80/20 containing 0.1% TEA, then adichloromethane/acetone mixture 50/50 containing 0.1% TEA.

34.0 mg of expected compound, of molecular formula C₃₄H₃₃N₂O₇SP(M=644.689 g) is recovered in the form of a yellow oil, i.e. a yield of23%.

1H NMR

In CDCl3, at 400 MHz, chemical shifts of the peaks in ppm andmultiplicity:

2.00 (m) and 2.48 (m): CH ₂—CH—C═O; 2.72 (d) and 3.12 (s): CH—CH ₂—N,3.75 (m): CH₂—CH—C═O₂; 4.71 (s) CH—CH₂—N, 5.18 [AB] CH ₂—C₆H₅; 7.35 (m):CH₂—C₆ H ₅ and 2.29 (m): CH ₃—CH═CH; 6.62 and 7.21 CH₃—CH═CH; 7.60-7.85P(C₆H₅)₃

MS (negative and positive electrospray) m/z:

[Manion]⁻=341

[Mcation]⁺=303

Example 11 1-propenyltriphenylphosphonium salt of trans methyl2-oxo-3-(sulphooxy)-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate

The operation is carried out as in Example 10, but starting from 207 mgof cis 1-(1,1-dimethylethyl) and 2-methyl4-hydroxy-1,2-pyrrolidinedicarboxylate.

In this way 12 mg of desired product of formula C₇H₁₀N₂O₇S (M=266.231 g)is obtained.

MS (negative and positive electrospray) m/z:

[Manion]⁻=265

[Mcation]⁺=303

Example 12a trans diphenylmethyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-3-carboxylate

8 ml of dichloromethane and 347 mg (1 mmole) of the hydrochloride of cisdiphenylmethyl 5-hydroxy-3-piperidinecarboxylate (described in ActaChem. Scand. Ser. B 35(4) 289-294) are mixed under an inert atmosphere.

The reaction medium is cooled down to 0° C., then 346 μl of TEA and 72μl of diphosgene are added.

The reaction medium is left to react for 15 minutes whilst maintainingthe temperature at 0° C., then the solvent is evaporated off underreduced pressure, followed by taking up in 25 ml of dry toluene andfiltering to eliminate the hydrochloride from the TEA.

553 μl of TEA is added to the filtrate and heating is carried out underreflux for 4 hours, followed by diluting with ethyl acetate, washingwith an aqueous solution containing 10% tartaric acid, then with asaturated aqueous solution of sodium chloride and drying the organicphase over magnesium sulphate.

The reaction medium is then evaporated under reduced pressure and 339 mgof crude product is recovered which is purified by chromatography onsilica, eluting with a toluene/ethyl acetate mixture 70/30.

In this way 146 mg of expected compound (M=337.378 g) is recovered,which corresponds to a yield of 43%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

2.15 (ddd) and 2.73 (dq): N—CH ₂—CHO—CH ₂; 2.92 (tt): O₂C—CH—; 3.00 (d)and 3.45 (d): N—CH ₂—CHO; 3.48 (dd) and 4.07 (dd): N—CH ₂—CH—CO₂; 4.79(dt): N—CH₂—CHO; 6.90 (s): CO₂—CH—(C₆H₅)₂; 7.33 (m): (C₆ H ₅)₂.

IR (CHCl₃): 1792, 1734; 1600, 1585, 1497 cm⁻¹.

MS (EI) m/z: [M]⁺=337, 292, 183, 167.

Example 12b trans-7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-3-carboxylicacid

320 mg of the compound obtained in Example 12a, 17 ml of acetone and 70mg of Pd/C catalyst at 20% by weight are mixed together.

Agitation is carried out under a hydrogen atmosphere at normal pressure.

After 2 hours 30 minutes, another 70 mg of catalyst is added and thereaction medium is left to react for another 1 hour 30 minutes, followedby filtering.

The solvent is evaporated off under reduced pressure and in this way 350mg of the crude product is obtained which is crystallized from pentane.

Filtration is carried out and in this way 158 mg of the sought productof molecular formula C₇H₉NO₄ (M=171.154 g) is recovered in the form of agrey solid. The corresponding yield is 89%.

1H NMR

In DMSO, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

2.10 (ddd) and 2.43 (dm): N—CH₂—CHO—CH ₂; 2.83 (tt): O₂C—CH—; 3.13 (d)and 3.27 (dm): N—CH₂—CHO; 3.40 (dd) and 3.72 (d): N—CH ₂—CH—CO₂H; 4.81(m): N—CH₂—CHO; 12.54 (broad s): CO₂ H.

IR (nujol): 1782, 1692 cm⁻¹.

MS (EI) m/z: [M]⁺=177, 155, 127, 82, 70.

Example 12c trans(4-nitrophenyl)methyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-3-carboxylate

30 mg (0.175 mmole) of the acid obtained in Example 12b and 0.5 ml ofdichloromethane are mixed together under an inert atmosphere. Then 26.8mg of 4-nitrobenzyl alcohol, 2.2 mg of DMAP and 37 mg of EDCI are added.

The reaction medium is left to react whilst agitating for 2 hours atambient temperature.

The organic phase is then diluted with dichloromethane, washed with a10% aqueous solution of tartaric acid and with a solution of phosphatebuffer at pH 7.

After drying the organic phase over sodium sulphate, and evaporating thesolvent under reduced pressure, 57 mg of crude product is obtained whichis purified by chromatography on silica eluting with a toluene/ethylacetate mixture 85/15.

The product is then crystallized from a mixture of ethyl ether andpentane in order to produce 34 mg of white crystals of the soughtcompound (M=306.277 g). The corresponding yield is 63.5%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

2.14 (ddd) and 2.84 (dm): N—CH₂—CHO—CHH₂; 2.90 (tt): O₂C—CH—; 3.10 and3.49 (dm); N—CH ₂—CHO; 3.43 (dd) and 4.14 (bd): N—CH ₂—CH—CO₂; 5.27[AB]: CO₂—CH ₂—C₆H₅; 7.56 and 8.24 [AA′BB′]: C—C₆H₅—NO₂.

IR (CHCl₃): 1799, 1789, 1741; 1609, 1526, 1495 cm⁻¹.

MS (EI) m/z: [M]⁺: 306, 170, 136, 126, 106, 82.

Example 13 6-(phenylmethyl)-1,6-diazabicyclo[3.2.1]octan-7-one Stage A

30.7 ml of TEA is added at about 0-5° C. to a solution of 20.71 g of3-amino-pyridine in 200 ml of methylene chloride. 25.5 ml of benzoylchloride is then added dropwise over 15 minutes and the reaction mediumis left to return to ambient temperature. Agitation is carried out for 1hour, followed by washing with water, then with a saturated solution ofsodium bicarbonate, the organic phase is dried over sodium sulphate andthe solvent is evaporated off under reduced pressure. 42.29 g ofexpected crystallized product (M=198.226 g) is obtained.

Stage B

4.3 ml of concentrated hydrochloric acid and 500 mg of rhodium onalumina at 5% by weight are added to a solution of 10 g of the productobtained in Stage A in 200 ml of methanol. The reaction medium is placeunder a hydrogen atmosphere at a pressure of 60-110 bars for 15 hours.

The reaction mixture is filtered, rinsed with methanol then the filtrateis concentrated under reduced pressure. The hydrochloride of theexpected product is obtained in a mixture with 10% of the hydrochlorideof the starting product.

The product is taken up in 250 ml of methylene chloride and 1.1equivalent of 1N soda is added. After agitation for 15 minutes, themethylene chloride is decanted, the organic phase is washed with water,followed by drying and evaporating under reduced pressure. The residueis chromatographed on silica eluting with a methylenechloride-methanol-triethylamine mixture 92/8/3.

7.4 g of expected crystallized product is obtained, i.e. a yield of 72%.

Stage C: N-(phenylmethyl)-3-piperidinamine

20 g of the product obtained as described in Stage B is dissolved in 600ml of 1,2-dimethoxyethane. 14.86 g of lithium aluminium hydride is addedto the solution over 30 minutes, followed by heating under agitation andunder an inert gas at 75-80° C. for 16 hours then cooling down to 0° C.and 11 ml of water is added over 45 minutes, without exceeding 12° C.Agitation is carried out for 10 minutes, followed by filtering andwashing the precipitate with methylene chloride. The filtrate isconcentrated under reduced pressure. 17.8 g of expected product isobtained in the form of an oil which is distilled under reduced pressure(boiling temperature: 114-121° C./0.8 mbar). 16 g of expected product isrecovered, i.e. a yield of 86%.

Stage D: 6-(phenylmethyl)-1,6-diazabicyclo[3.2.1]octan-7-one

1.06 g of product obtained in Stage C is dissolved in 28 cm³ of toluene,then the reaction medium is cooled down to 0° C. and 337 μl ofdiphosgene is added under an inert gas. The temperature is allowed torise and is maintained at 20° C. for 2 hours. Concentration is carriedout under reduced pressure then the residue is chromatographed on silicaeluting successively with methylene chloride-acetone 95/5 then 80/20 andfinally methylene chloride-methanol, triethylamine 92/8/3 and 362 mg ofexpected product C₁₃H₁₆N₂O (M=216.85 g) is obtained i.e. a yield of 30%.

VPC/Mass spectrum (EI) m/z: [M]⁺=216, 125, 91.

IR (CHCl₃): 1718; 1498 cm⁻¹.

Example 14 6-benzoyl-1,6-diazabicyclo[3.2.1]octan-7-one Stage A:3-(benzylamino)-1-piperidinecarboxylic

5 g of product obtained in Stage B of Example 13, is dissolved in 1.25 lof anhydrous toluene under a nitrogen atmosphere then 3.4 ml of TEA isadded and 1.47 ml of diphosgene is introduced at 0-5° C. over 3 minutes.After 20 minutes at 0-5° C., the reaction medium is left to heat up to20° C., is maintained under agitation for 75 minutes, then the solventis evaporated off under reduced pressure. The residue is chromatographedon silica eluting with a methylene chloride-acetone mixture 8/2. 3.44 gof expected product is obtained (yield of 52.6%).

Stage B 6-benzoyl-1,6-diazabicyclo[3.2.1]octan-7-one

48 mg of sodium hydride at 50% in dispersion in oil and 20 ml of THF areintroduced under a nitrogen atmosphere. The reaction medium is cooleddown to about 0-5° C., then 266 mg of the product obtained in Stage A isadded in one go.

The temperature is allowed to rise to ambient temperature, then 60 μl ofacetic acid and 10 ml of phosphate buffer at pH 7 are added.

Then a little ethyl acetate is added followed by decanting andreextracting with ethyl acetate. The organic phase is dried overmagnesium sulphate, then the solvents are evaporated off under reducedpressure.

The crude product is chromatographed on silica eluting withdichloromethane containing 2% acetone.

In this way 143 mg of the sought product C₁₃H₁₂N₂O₂ (M: 228.25 g) isobtained. The corresponding yield is 62%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.20-2.15 (m) and 2.42 (m): NCH—CH ₂—CH ₂—; 2.80 (d) —2.93 (d); 3.11(m); 3.28 to 3.58 (m): CH ₂—N, 4.54 (m): CH—N, 7.43 (m); 7.55 (m); 7.69(m): C₆ H ₅

IR (CHCl₃): 1758, 1672; 1605, 1586, 1492;

MS (EI) m/z: [M]⁺=230, 125, 105, 77

Example 15 7-oxo-1,6-diazabicyclo[3.2.1]octan-6-acetic acid Stage A5-[(1,1-dimethylethyl)dimethylsilyl]-1,6-diazabicyclo[3-2-1]octan-7-one]

843 mg of lithium is placed under a nitrogen atmosphere and condensed at−70° C. with 320 ml of ammonia. 7.56 g (34.8 mmoles) of the productobtained in Example 13 in 160 ml of tetrahydrofuran is added at −70° C.over 10 minutes. Agitation is carried out for 5 minutes then the ammoniais distilled under a stream of nitrogen whilst heating slowly to 20° C.7.9 g of (1,1-dimethylethyl) dimethylsilyl chloride in 10 cm³ oftetrahydrofuran is added slowly to the suspension obtained, at 20° C.,then maintained under agitation for 10 minutes. Then 160 cm³ of ethylacetate then 60 cm³ of a 10% aqueous solution of tartaric acid areadded, followed by decanting, reextracting with ethyl acetate washingthe organic phase with water, drying it over sodium sulphate andevaporating the solvent under reduced pressure. The oil obtained ischromatographed on silica with 10% water, eluting with methylenechloride then with a methylene chloride-acetone mixture 8/2 and 3.04 gof expected product is obtained (yield: 36.2%).

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

0.21 (S) and 0.40 (S): SiCH ₃; 0.97 (S): SitBu; 1.5 to 1.8 (m) and 2.07(m): N—CH—CH ₂—CH ₂; 2.85 (d) and 3.32 (m); —CH—CH ₂—N, 2.93 (dt) and3.32 (m): —CH₂—CH ₂—N, 3.65 (m): CH—N.

IR (CHCl₃): 1710; 842 cm⁻¹

MS (EI) m/z: [M]⁺: 240, 225, 183, 100, 83, 57.

Stage B phenylmethyl 7-oxo-1,6-diazabicyclo[3-2-1]octan-6-acetate

1.44 g (5.99 mmoles) of the product obtained in Stage A is dissolvedunder a nitrogen atmosphere in 14.4 ml of tetrahydrofuran then 941 μl ofphenylmethyl bromoacetate is added and then, 6 ml of a 1 M solution oftetra-n-butyl ammonium fluoride in tetrahydrofuran is added dropwise.Agitation is carried out for 10 minutes at 20° C. then the reactionmedium is diluted with 15 ml of ethyl acetate and 5 ml of an aqueoussolution of phosphate buffer at pH=7 is added, followed by decanting,reextracting with ethyl acetate, washing the organic phase with water,drying it over sodium sulphate and the solvent is evaporated off underreduced pressure. The oily residue is chromatographed on silica with 10%water eluting with a methylene chloride-acetone mixture 8/2. 140 mg ofthe expected product is obtained. The corresponding yield is 9%.

IR (CHCl₃): 1746, 1720 cm⁻¹.

MS (EI) m/z: [M]⁺=274, 183, 155, 139, 91, 83.

Stage C: 7-oxo-1,6-diazabicyclo[3.2.1]octane-6-acetic acid

137 mg of the product obtained in Stage B is dissolved in 1.5 ml ofethyl acetate, then 14 mg of 10% palladium on carbon is added to thesolution and the reaction medium is placed under a hydrogen atmosphere.After 15 minutes another 15 mg of palladium on carbon is added and thereaction medium is maintained under agitation for 15 minutes. Thecatalyst is filtered, followed by rinsing with ethyl acetate, then withacetone and with methanol and the solvent is evaporated off underreduced pressure. A total of 68 mg of crude product is obtained which iscrystallized from ether. 58 mg of the expected product of molecularformula C₁₅H₁₈N₂O₃ (M=274.321 g) is obtained. The corresponding yield is63%.

1H NMR

In CDCl3, at 400 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.48 (m), 1.63 (m), 1.73 (m) and 1.86 (m): N—CH—CH ₂—CH ₂; 2.85 to 3.00(m), 3.14 (dm) and 3.64 (m): CH ₂—N—CH ₂ and CH—N, 3.78 and 4.14 [AB]:CON—CH ₂—CO.

MS (EI) m/z: [M]⁺=184, 139, 125, 111, 97, 83.

Example 16 7-oxo-N-phenyl-1,6-diazabicyclo[3.2.1]octane-6-carboxamide

1 ml of tetrahydrofuran and 99 mg (0.41 mmole) of the compound obtainedin Stage A of Example 15 are mixed under an inert gas.

50 μl of phenyl isocyanate then 450 μl of a 1M solution oftetrabutylammonium fluoride in THF are added successively.

The reaction medium is left to react for 10 minutes, then diluted withethyl acetate, followed by washing with water, decanting and drying theorganic phase over magnesium sulphate. The solvent is evaporated offunder reduced pressure. In this way 140 mg of crude product is obtainedwhich is purified by chromatography on silica eluting with adichloromethane/ethyl acetate mixture 90/10.

21 mg of the title compound, of molecular formula C₁₃H₁₅N₃O (M=245.283g) is recovered which corresponds to a yield of 20%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.78 (m), 2.02 (m) and 2.17 (m): N—CH—CH ₂—CH ₂; 2.88 (d), 3.13 (dt) and3.42 (m): CH ₂—N—CH ₂; 4.49 (m): CH—N, 7.11 (t); 7.34 (t) and 7.54 (d):C₆ H ₅; 10.05: NH.

IR (CHCl₃): 3302, 3266; 1734; 1700; 1602, 1553, 1501 cm⁻¹.

MS (EI) m/z: [M]⁺: 245, 153, 126, 119, 98, 92.

Example 17a6-[1-(phenylmethyl)-1H-tetrazol-5-yl]-1,6-diazabicyclo[3.2.1]octan-7-one

480 mg (2 mmoles) of the compound obtained in Stage A of Example 15 isplaced under an inert gas.

Then a solution of 712 mg of 5-fluoro-1-(phenylmethyl)-1H-tetrazole in1.5 ml of tetrahydrofuran and then 2 ml of a 1 M solution oftetrabutylammonium fluoride in THF is added. The reaction medium is leftto react for 1 minute, followed by diluting with ethyl acetate, washingwith water, decanting, drying the organic phase over magnesium sulphateand the solvent is evaporated off under reduced pressure.

1.06 g of an oily product is obtained which is chromatographed on silicain a dichloromethane/ethyl acetate mixture 90/10.

In this way 143 mg of expected compound of molecular formula C₁₄H₁₆N₆O(M=284.324 g) is obtained in the form of an amorphous white product. Thecorresponding yield is 25%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.80 (m), 2.04 (m) and 2.67 (m): N—CH—CH ₂—CH ₂; 2.83 (d), 2.85 (dm),3.10 (dd) and 3.44 (dd): CH ₂—N—CH ₂; 3.99 (m): CH—N, 5.63 and 5.88[AB]: C₆H₅—CH ₂; 7.18 (m) and 7.32 (m): C₆ H ₅.

Example 17b 6-(1H-tetrazol-5-yl)-1,6-diazabicyclo[3.2.1]octan-7-one

120 mg of the product obtained in Example 17a and 2.4 ml of amethanol/ethyl acetate mixture 90/10 are mixed together then 2.4 ml ofTHF is added until total dissolution is obtained.

Then 24 mg of 10% palladium catalyst on carbon is added then agitationis carried out under a hydrogen atmosphere. After reaction for 3 hours,the catalyst is filtered, followed by rinsing with atetrahydrofuran/methanol mixture, then the solvent is evaporated offunder reduced pressure. Then the product crystallizes from ethyl ether.

In this way 72 mg of the title compound of molecular formula C₇H₁₀N₆O(M=194.198 g) is obtained in the form of a white crystallized product.The corresponding yield is 88%.

1H NMR

In DMSO, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.63 (m), 1.89 (m) and 2.07 (m): N—CH—CH ₂—CH ₂; 3.14 to 3.20 (m) and3.43 (m): CH ₂—N—CH ₂; 4.51 (m): CH—N.

IR (Nujol): 1744; 1594 cm⁻¹.

MS (EI) m/z: [M]⁺=194, 165, 124, 111, 98, 83, 68, 56, 41.

Example 18 6-acetyl-1,6-diazabicyclo[3.2.1]octan-7-one

140 mg (0.582 mmoles) of the compound obtained in Stage A of Example 15is dissolved in 1.4 ml of THF.

55 μl of acetic anhydride then 0.58 ml of a 1 M solution oftetrabutylammonium fluoride in THF are added successively to thesolution obtained, followed by diluting with ethyl acetate, washing withwater, decanting, drying the organic phase over magnesium sulphate, thenthe solvent is evaporated off under reduced pressure.

In this way 116 mg of a crude oil is obtained which is chromatographedon silica with a dichloromethane/acetone mixture 80/20.

In this way 18 mg of expected compound, of molecular formula C₈H₁₂N₂O₂(M=168.196 g) is obtained, which corresponds to a yield of 18%.

1H NMR

In DMSO, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.65 to 2.20 (m): N—CH—CH ₂—CH ₂; 2.54 (s): CH ₃CO—N, 2.83 (d), 3.33(dm), 3.10 (m) and 3.45 (dd) CH ₂—N—CH ₂; 4.55 (m):

O═C—N—CH.

IR (CHCl₃): 1758, 1696 cm⁻¹.

MS (EI) m/z: [M]⁺=168, 140, 126, 98, 43.

Example 19a 6-(phenylmethoxy)-1,6-diazabicyclo[3.2.1]octan-7-one

44.02 g (0.22 mole) of 1,1-dimethylethyl 3-oxo-1-piperidinecarboxylate(C₁₀H₁₇NO₃, M=199.251 g) (described in J. Med. Chem. 1986, 29, 224-229)is dissolved in 440 ml of ethanol.

Then 38.79 g of O-benzyl-hydroxylamine hydrochloride is added. 54 ml ofpyridine is then introduced dropwise, into the suspension.

The reaction medium is left to react whilst agitating for 4 hours atapproximately 25° C., then the solvent is evaporated off under reducedpressure, followed by taking up in a mixture of dichloromethane andethyl acetate, filtering and rinsing with dichloromethane, then with amixture of dichloromethane and ethyl acetate. The filtrate is thenconcentrated to dryness under reduced pressure.

In this way 69.8 g of a light yellow oil is obtained which is purifiedby chromatography on silica. The eluent used is a cyclohexane/ethylacetate mixture 80/20.

57.21 g of 1,1-dimethylethyl3-[(phenylmethoxy)imino]-1-piperidinecarboxylate, of molecular formulaC₁₇H₂₄N₂O₃ (M=304.39 g) is recovered, in the form of a very pale yellowoil. The corresponding yield is 85%.

24.82 g (0.0815 mmole) of the oxime obtained previously is dissolved in163 ml of ethanol cooled down to −10° C. under nitrogen. Then 25 ml of aborane-pyridine complex is added then, 204 ml of 2N hydrochloric acid isadded dropwise over one hour 15 minutes. The solution is agitated for 1hour 15 minutes at −5° C., then treated with 100 ml of a saturatedsolution of sodium hydrogen carbonate, then with 35 g of sodiumcarbonate, which are add by small portions. The pH is then 7-8.

The reaction medium is extracted with ethyl acetate.

The organic phases are combined, dried over sodium sulphate and thesolvent is evaporated off under reduced pressure. In this way 39.0 g ofa colourless oily liquid is obtained which is taken up in 400 ml ethylacetate.

The solution is washed with a 0.05 N aqueous solution of hydrochloricacid, then the phases organic are combined and the solvent is evaporatedoff under reduced pressure.

35.5 g of a colourless oily liquid is recovered which is purified bychromatography on silica, eluting with a dichloromethane/ethyl acetatemixture 95/5, then with a dichloromethane/ethyl acetate mixture 80/20.

In this way 17.89 of 1,1-dimethylethyl3-[(phenylmethoxy)amino]-1-piperidinecarboxylate of molecular formulaC₁₇H₂₆N₂O₃ (M=306.41 g), is recovered in the form of a colourless oil.The corresponding yield is 72%.

6.72 g (21.9 mmoles) of the piperidine obtained previously is dissolvedin 22 ml of ethyl acetate cooled down to −10° C. 28 ml of a 4.0 mol/lsolution of anhydrous hydrochloric acid in ethyl acetate is addeddropwise, over 30 minutes.

After 1 hour at 0° C., 40 ml of ethyl ether is added, thedihydrochloride precipitate is filtered and washed with ethyl ether.

In this way 3.87 g of a white solid is obtained.

By crystallizing the filtrate, 1.80 g of the desired product is alsoobtained.

The product obtained is taken up in 60 ml of 1 N soda and 120 ml ofethyl acetate. After decanting, the aqueous phase is saturated withsodium chloride, then extracted twice with ethyl acetate. The organicphases are combined and dried over magnesium sulphate then concentratedto dryness under reduced pressure.

In this way 3.67 g of N-(phenylmethoxy)-3-piperidinamine, of molecularformula C₁₂H₁₈N₂O (M=206.29 g) is obtained, which corresponds to a yieldof 81%.

518 mg (2.5 mmoles) of the compound obtained previously is dissolved in5 ml of anhydrous dichloromethane, then 0.5 ml of TEA is added.

The whitish suspension obtained is cooled down to −65° C., then 12.5 mlof a 0.10 mol/l solution of diphosgene in dichloromethane is added over15 minutes.

After reaction for 45 minutes, the colourless solution is diluted with15 ml of dichloromethane and treated with 15 ml of water.

The medium is left to settle, then the aqueous phase is extracted with20 ml of dichloromethane.

The combined organic phases are dried over magnesium sulphate, thenconcentrated to dryness under reduced pressure. In this way a paleyellow oil is obtained which is purified by chromatography on silicaeluting with an ethyl acetate mixture 90/10, then adichloromethane/ethyl acetate mixture 80/20.

In this way 196 mg of expected compound of molecular formula C₁₃H₁₆N₂O₂,(M=232.28 g) is recovered in the form of a colourless oil. Thecorresponding yield is 34%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.59 (m) and 1.93 to 2.18 (m): N—CH—CH ₂—CH ₂; 2.73 (dt), 2.94 (dt),3.17 (dt) and 3.40 (dd): CH ₂—N—CH ₂; 3.29 (t) N—CH; 4.89 (d): N—O—CH ₂—(C₆H₅); 7.38: C₆ H ₅.

IR (CHCl₃): 1747; 1498 cm⁻¹.

MS (EI) m/z: [M]⁺=232, 91.

Example 19b 6-(acetyloxy)-1,6-diazabicyclo[3.2.1]octan-7-one

95 mg (0.41 mmole) of the compound obtained in Example 19a is dissolvedin 5 ml of methanol, agitation is carried out with 8 mg of palladium oncarbon at 10% by weight, then the suspension is placed under a hydrogenatmosphere under normal pressure for 1 hour at 25° C., then the catalystis filtered.

After evaporation of the solvent under reduced pressure, 70 mg of whitecrystals is obtained.

The crystals are taken up in 2 ml of anhydrous dichloromethane. Thesolution is cooled down to −10° C. under nitrogen. Then 70 μl ofpyridine then 40 μl of acetic anhydride are added and agitation iscarried out for 20 minutes. Concentration is carried out under reducedpressure and 75 mg of white crystals are obtained which are purified onsilica, eluting with a dichloromethane ethyl acetate mixture 80/20.

49 mg of expected compound (M=184.20 g) is recovered in the form of awhite solid. The corresponding yield is 65%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.60 to 2.2: N—CH—CH ₂—CH ₂; 2.24 (s): CH₃; 2.95 (d) and 3.54 (dm): N—CH₂—CH; 3.07 (dt) and 3.54 (bdd): N—CH ₂—CH₂; 3.94 (bt): O═C—N—CH.

IR (CHCl₃): 1798; 1764 cm⁻¹.

MS (EI) m/z: [M]⁺=184, 142, 125, 43.

Example 19c 6-(benzoyloxy)-1,6-diazabicyclo[3.2.1]octan-7-one

The operation is carried out in a similar manner to that which has beendescribed in Example 19b starting from 205 mg of the compound preparedin Example 19a and 200 mg of benzoic anhydride.

In this way 64 mg of expected compound of molecular formula C₁₃H₁₄N₂O₃(M=246.27 g) is obtained i.e. a yield 30%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.64 to 1.95 (m) and 2.10 to 2.35 (m): CH—CH ₂—CH ₂; 3.02 (d) and 3.65(dm): N—CH ₂—CH; 3.13 (dt) and 3.55 (bdd): N—CH ₂—CH₂; 4.09 (bt):O═C—N—CH; 7.49 (m): 7.65 (tt); 8.12 (m): C₆ H ₅.

IR (CHCl₃): 1774, 1756; 1602, 1585, 1495 cm⁻¹.

MS (EI) m/z: [M]⁺=246, 105, 77.

Example 19d 6-(1-oxopropoxy)-1,6-diazabicyclo[3.2.1]octane-7-one

The operation is carried out in a similar manner to that which has beendescribed in Example 19c, starting from 163 mg of the compound preparedin Example 19a and 70 μl of propionyl chloride.

In this way 17 mg of expected compound of molecular formula C₉H₁₄N₂O₃(M=198.23 g) is obtained, i.e. a yield of 12%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.25 (t): O═C—CH₂—CH ₃; 1.65 (m), 1.78 (m) and 2.10 (m): N—CH—CH ₂—CH ₂;2.52 (m) O═C—CH ₂—CH₃; 2.94 (d) and 3.55 (bd): N—CH ₂—CH; 3.07 (dt) and3.48 (dd): N—CH ₂—CH₂; 3.93 (m): N—CH₂—CH.

IR (CHCl₃): 1792; 1763 cm⁻¹.

MS (EI) m/z: [M]⁺=198, 170, 142, 125, 97, 57.

Example 19e6-[[(4-methylphenyl)sulphonyl]oxy]-1,6-diazabicyclo[3.2.1]octan-7-one

The operation is carried out in a similar manner to that which has beendescribed in Example 19d, starting from 139 mg of the compound preparedin Example 19a and 126 mg of tosyl chloride.

In this way 77 mg of expected compound of molecular formula C₁₃H₁₆N₂O₄S(M=296.35 g) is obtained i.e. a yield of 44%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.55 and 2.99 (m): N—CH—CH ₂—CH ₂; 2.45 (s): CH₃; 2.89 (d), 3.00 (dt),3.29 (dt) and 3.39 (dd): CH ₂—N—CH ₂; 4.04 (m): N—CH; 7.35 and 7.91[AA′BB′] CH₃—C₆ H ₄—SO₂.

IR (CHCl₃): 1775; 1599, 1495, 1383; 1193, 1180 cm⁻¹.

MS (EI) m/z: [M]⁺=296, 155, 141, 125, 91.

Example 19f 6-[(methylsulphonyl)oxy]-1,6-diazabicyclo[3.2.1]octan-7-one

The operation is carried out in a similar manner to that which has beendescribed in Example 19e starting from 211 mg of the compound preparedin Stage 19a and 80 μl of mesyl chloride.

In this way 50 mg of expected compound of molecular formula C₁₇H₁₂N₂O₄S(M=220.25 g) is obtained i.e. a yield of 25%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.56 and 2.38 (m): N—CH—CH ₂—CH ₂; 3.00 (d), 3.12 (dt) and 3.49 (m):N—(CH ₂)₂; 3.26 (s): CH₃; 4.12 (m): N—CH.

IR (CHCl₃): 1775; 1381, 1187 cm⁻¹.

MS (EI) m/z: [M]⁺=220, 141, 125, 97, 79.

Example 19 g6-[(4-nitrophenyl)sulphonyl]oxy]-1,6-diazabicyclo[3.2.1]octan-7-one

The operation is carried out in a similar manner to that which has beendescribed in Example 19f starting from 270 mg of the compound preparedin Example 19a and 283 mg of 4-nitrobenzenesulphonyl chloride.

In this way 205.5 mg of expected compound of molecular formulaC₁₂H₁₃N₃O₆S (M=327.32 g) is obtained i.e. a yield of 54%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.64 (dt), 1.84 (m), 1.99 (m), 2.31 (dm): NCH—CH ₂—CH ₂; 2.94 (d), 3.30(dt), 3.04 (dt), 3.40 (bdd): N(CH ₂)₂; 4.14: O═C—N—CH; 8.25 and 8.41[AA′BB′]: NO₂—C₆ H ₄SO₂.

IR (CHCl₃): 1776; 1610, 1590, 1538; 1393, 1191 cm⁻¹.

MS (EI) m/z: [M]⁺=327, 186, 141, 125, 111.

Example 206-[[(4-methylphenyl)sulphonyl]amino]-1,6-diazabicyclo[3.2.1]octan-7-one

5 g (25.1 mmole) of 1,1-dimethylethyl 3-oxo-1-piperidinecarboxylate(described in J. Med. Chem. 1986, 29, 224-229) (C₁₀H₁₇NO₃, M=199.251 g)is dissolved in 50 ml of dichloromethane.

4.67 of tosylhydrazine is then added to the solution which is left toreact for 2 hours under agitation, then the solvent is evaporated offunder reduced pressure.

In this way, 9.56 g of 1,1-dimethylethyl3-[2-[(4-methylphenyl)sulphonyl]hydrazono]-1-piperidinecarboxylate, ofmolecular formula C₁₇H₂₅N₃O₄S (M=367.47 g) is obtained with aquantitative yield.

4.5 g of the compound obtained previously (12.2 mmoles), 90 ml of amethanol/tetrahydrofuran mixture 50/50, and a few grains of bromocresolgreen are mixed together under an inert gas.

Then 1.62 g of NaBH₃CN is added, followed by cooling down to about 0-5°C., and a 0.7 mol/l solution of gaseous hydrogen chloride in methanol isintroduced, in such a way so as to maintain the pH of the medium between3.8 and 5.4.

The reaction medium is left to react whilst agitating for 2 hours and 30minutes.

2/3 of the solvents are evaporated under reduced pressure, then 200 mlof dichloromethane is added followed by washing with a saturated aqueoussolution of sodium bicarbonate.

The organic phase is dried over sodium sulphate and the solvent isevaporated off under reduced pressure.

In this way 4.48 g of 1,1-dimethylethyl 3-[2-[(4-methylphenyl)sulphonyl]hydrazino]-1-piperidinecarboxylate of molecular formulaC₁₇H₂₇N₃O₄S (M=369.486 g) is obtained.

The corresponding yield is 99%.

4.48 g of the compound obtained previously and 9 ml of ethyl acetate aremixed under an inert gas at 0° C.

30 ml of a 4 mol/l solution of gaseous hydrogen chloride in ethylacetate is added, agitation is carried out for 15 minutes followed byfiltering and washing the hydrochloride with ethyl acetate. After dryingunder reduced pressure, 3.48 g of the dihydrochloride of2-(3-piperidinyl)hydrazide of 4-methyl-benzenesulphonic acid, ofmolecular formula C₁₂H₁₉N₃O₂S, 2HCl (M=342.289 g) is obtained. Thecorresponding yield is 84%.

Then 3.48 g of the compound obtained previously is dissolved in 5 ml ofdemineralized water. 10.2 ml of a 2N aqueous solution of soda is addedunder vigorous agitation.

A precipitate forms after 1 to 2 minutes of contact. Agitation is thencarried out for 10 minutes, then the precipitate is filtered and washedwith water, then with ethyl acetate.

The solid obtained is dried under reduced pressure.

In this way 2.21 g of 2-(3-piperidinyl)hydrazide of4-methyl-benzenesulphonic acid, of molecular formula C₁₂H₁₉N₃O₂S(M=269.328 g) is obtained. The corresponding yield is 81%.

500 mg (1.85 mmole) of the amine obtained previously and 20 ml oftetrahydrofuran are mixed under an inert gas.

112 μl of diphosgene then 517 μl of TEA and 23 mg of DMAP are added, ata temperature comprised between 0 and 5° C., to the suspension obtained.

The reaction medium is left to react whilst agitating and whilstallowing the temperature to rise to 20° C., followed by diluting withethyl acetate, then washing with a 10% aqueous solution of tartaricacid, then with demineralized water.

The organic phase is dried over magnesium sulphate, then the solvent isevaporated off under reduced pressure.

769 mg of a crude product is obtained which is dissolved in 7 ml ofdichloromethane and 517 μl of TEA.

The reaction medium is left to react overnight under agitation, followedby diluting with dichloromethane, washing with water, drying over sodiumsulphate and the solvent is evaporated off under reduced pressure.

The foam obtained (395 mg) is purified by chromatography on silica witha dichloromethane/ethyl acetate mixture 80/20.

44 mg of expected compound, of molecular formula C₁₃H₁₇N₃O₂S (M=295.362g) is recovered. The corresponding yield is 8%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.55 to 1.80 (m) and 2.18 (m): N—CH—CH ₂—CH ₂; 2.42 (s): CH₃; 2.88 (d)and 2.93 (m); N—CH ₂—CH; 3.18 to 3.32 (m): N—CH ₂—CH₂; 4.08 (m):N—CH—CH₂; 6.98 (bs): NH.

IR (CHCl₃): 3264, 1737, 1599, 1490 cm⁻¹.

MS (positive electrospray) m/z: [M+Na]⁺=318, [M+H]⁺=296

Example 216-[(4-methylphenyl)sulphonyl]-1,6-diazabicyclo[3.2.1]octan-7-one

305 mg (1.52 mmole) of 1,1-dimethylethyl 3-amino-1-piperidinecarboxylate(described in J. Med. Chem. 1992, 35, 4334-4343), of molecular formulaC₁₀H₂₀N₂O₂ (M=200.282 g) is dissolved in 3 ml of anhydrousdichloromethane.

Then, 212 μl of TEA is added, followed by cooling down to 5° C. and 278mg of tosyl chloride is added. Agitation is carried out whilst allowingthe temperature to return to 20° C. and the reaction medium is left toreact for 2 hours, followed by diluting with dichloromethane and washingfirstly with a 10% aqueous solution of tartaric acid then with asolution of phosphate buffer at pH=7.

After separating, the organic phase is dried over magnesium sulphate,then the solvent is evaporated off under reduced pressure. In this wayan oil is obtained which is purified by chromatography on silica elutingwith a dichloromethane/ethyl acetate mixture 9/1.

440 mg of 1,1-dimethylethyl3-[[(4-methylphenyl)sulphonyl]amino]-1-piperidinecarboxylate (describedin J. Med. Chem. 1992, 35, 4334-4343) of molecular formula C₁₇H₂₆N₂O₄S(M=354.472 g) is recovered. The corresponding yield is 82%.

A mixture of 425 mg of the compound obtained previously and 2.1 ml of atrifluoroacetic acid/dichloromethane mixture 50/50 is cooled down to0-5°.

The reaction medium is kept under agitation at 5° C. for 30 minutes.

Then the solvent is evaporated off under reduced pressure in order toobtain 403 mg of 4-methyl-N-(3-piperidinyl)-benzenesulphonamidetrifluoroacetate of molecular formula C₁₄H₁₉F₃N₂O₄S (M=368.377 g).

228 mg of the compound obtained previously is suspended in 2 ml ofmethanol, then treated with an excess of DOWEX 21K 20-50 Mesh resinactivated with soda, followed by filtering, rinsing the resin withmethanol, then the filtrate is evaporated under reduced pressure.

In this way 123 mg of 4-methyl-N-(3-piperidinyl)-benzenesulphonamide ofmolecular formula C₁₂H₁₈N₂O₂S (M=254.353 g) is recovered.

118 mg of the amine obtained previously is dissolved under an inert gasin 1.2 ml of dichloromethane.

Then 98 μl of TEA and then 28 μl of diphosgene are introducedsuccessively. The reaction medium is left to react whilst agitating for30 minutes at 0-5° C., followed by diluting with dichloromethane,washing the organic phase with a 10% aqueous solution of tartaric acid,then with water. After drying over sodium sulphate, filtration andevaporation of the solvent under reduced pressure, the crude product ispurified by chromatography on silica eluting with adichloromethane/acetone mixture 95/5.

In this way 112 mg of the chloride of3-[[(4-methylphenyl)sulphonyl]amino]-1-piperidinecarboxylic acid, ofmolecular formula C₁₃H₁₇ClN₂O₃S (M=316.308 g) is obtained. Thecorresponding yield is 76%.

10 mg of sodium hydride (in suspension at 55-65% in the oil) and 2 ml ofanhydrous tetrahydrofuran are mixed together under an inert atmosphere.

Then 71 mg of the product obtained previously is added.

Agitation is carried out at ambient temperature for 15 minutes, then 12μl of acetic acid and 2 ml of solution of phosphate buffer at pH=7 areadded.

Agitation is carried out for another 5 minutes, then 5 ml ethyl acetateis added, followed by leaving to settle, then reextracting with ethylacetate. After separating, the organic phase is dried over magnesiumsulphate, filtered and the solvent is evaporated off under reducedpressure.

In this way 65 mg of crude product is obtained which is purified bychromatography on silica, eluting with a dichloromethane/acetone mixture95/5.

In this way 40 mg of expected compound, of molecular formula C₁₃H₁₆N₂O₃S(M=280.348 g) is recovered. The corresponding yield is 64%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity (presence of two conformers 90/10):

1.46 (m), 1.76 (m) and 2.08 (dm): NCH—CH ₂—CH ₂; 2.44 (s) and 2.45 (s):CH₃; 2.82 (d) and 2.98 (m) and 3.28 to 3.50 (m): —N—(CH ₂)₂; 4.55 (m)and 4.65 (m): CO—N—CH; 7.33 and 7.78, 7.35 and 8.02 [AA′BB′]CH₃—C₆H₄—SO₂.

IR (CHCl₃): 1758, 1598, 1995, 1367, 1169 cm⁻¹.

MS (EI) m/z: [M]⁺: 280, 216, 155, 125, 97, 91.

Example 22 6-oxa-1-azabicyclo[3.2.1]oct-3-en-7-one

5 ml of dichloromethane and 68 mg of 1,2,3,6-tetrahydro-pyridin-3-olhydrochloride (M=135.5 g) (described in Chem. Pharm. Bull.30(10)3617-3623 (1982)) are mixed together under an inert gas.

33 μl of diphosgene is added and agitation is carried out for 5 minutesat 0° C. Then 140 μl of TEA and 61 mg of DMAP are added.

The reaction medium is left to react at ambient temperature for 2 hours,followed by diluting with dichloromethane and washing with a 10% aqueoussolution of tartaric acid then with water, decanting and drying theorganic phase over magnesium sulphate. The solvent is evaporated offunder reduced pressure. In this way 5 mg of crude product is obtainedwhich is purified by chromatography on silica, eluting withdichloromethane then a dichloromethane/ethyl acetate mixture 95/5.

In this way 3 mg of expected compound, of molecular formula C₆H₇NO₂(M=125 g) is recovered. The corresponding yield is 5%.

Example 23 phenylmethyltrans-3-benzoyl-2-oxo-4-oxa-1,3-diazabicyclo[3.2.1]octane-7-carboxylate

5.50 g (13.7 mmoles) of cis 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-[(methylsulphonyl)oxy]-1,2-pyrrolidine dicarboxylate(described in J. Org. Chem. 1991, 56, 3009-3016), of molecular formulaC₁₈H₂₅NO₇S (M=399.466 g) and 110 ml of dimethylformamide are mixedtogether under an inert gas then 2.58 g of N-hydroxyphthalimide, then1.52 g of potassium hydrogen carbonate is added.

The reaction medium is heated under agitation at 100° C. and maintainedat this temperature for 4 hours.

The reaction medium is cooled down to 20° C., 220 ml of water and iceare added, then extraction is carried out with isopropyl ether, followedby drying over magnesium sulphate, then evaporating to dryness underreduced pressure.

The residue is chromatographed on silica, eluting with adichloromethane/ethyl acetate mixture 90/10.

In this way 3.06 g of trans 1-(1,1-dimethylethyl) and 2-(phenylmethyl4-[(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)oxy]-1,2-pyrrolidinedicarboxylate),of molecular formula C₂₅H₂₆N₂O₇ (M=466.494 g) is recovered Thecorresponding yield is 47%.

3.24 g (6.94 mmoles) of the phthalimide obtained previously is dissolvedin 33 ml of dichloromethane.

372 μl of hydrazine hydrate is added.

Agitation is again carried out for 2 hours 30 minutes at 20° C.

The precipitate formed is filtered, rinsed with dichloromethane, thenthe solvent is evaporated off under reduced pressure.

2.91 g of crude product is obtained which is purified by chromatographyon silica, eluting with a dichloromethane/ethyl acetate mixture 90/10,then 80/20 and 50/50.

In this way a total of 942 mg of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)4-(aminooxy)-1,2-pyrrolidinedicarboxylate, of molecularformula C₁₇H₂₄N₂O₅ (M=336.39 g) is recovered. The corresponding yield is40%.

853 mg of the compound obtained previously (2.53 mmoles) and 8.5 ml ofanhydrous dichloromethane are mixed together under an inert gas.

The reaction medium is cooled down to about 0-5° C., then 706 μl of TEAand 588 μl of benzoyl chloride are added.

Agitation is carried out for 10 minutes at 0-5° C., then the reactionmedium is left to heat up to 20° C. and left again to react for 30minutes.

The organic phase is washed with a 10% aqueous solution of tartaricacid, then with water, followed by decanting and drying the organicphase over sodium sulphate, the solvent is evaporated off under reducedpressure.

In this way 1.38 g of product is obtained, which is mixed with 25 ml ofdichloromethane. The reaction medium is cooled down to about 10-15° C.and 123 μl of hydrazine hydrate is added.

The reaction medium is left to react whilst agitating at 20° C. for twohours and 30 minutes.

The solvent is evaporated off under reduced pressure.

In this way 1.13 g of crude product is obtained which is purified bychromatography on silica, eluting with a dichloromethane/ethyl acetatemixture 80/20.

948 mg of trans 1-(1,1-dimethylethyl) and2-(phenylmethyl)-4-[(benzoylamino)oxy]-1,2-pyrrolidinedicarboxylate, ofmolecular formula C₂₄H₂₈N₂O₆ (M=440.50 g) is recovered.

The overall yield is therefore 85%.

948 mg of the compound obtained previously is dissolved under agitationin 2 ml ethyl acetate.

The reaction medium is cooled down to 0-5° C., then 4.7 ml of anapproximately 4.6 M solution of gaseous hydrogen chloride in ethylacetate is added in one go.

After 1 hour, the solvent is evaporated off under reduced pressure andthe product is taken up 3 times in ethyl ether.

The solvent is evaporated off under reduced pressure. In this way 842 mgof the hydrochloride of trans phenylmethyl4-[(benzoylamino)oxy]-2-pyrrolidine carboxylate, is obtained in the formof a white friable foam of formula C₁₉H₂₀N₂O₄, HCl (M=376.84 g).

The yield is quantitative.

47 mg (0.125 mmole) of the hydrochloride obtained previously isdissolved under an inert gas in 0.5 ml of dichloromethane. 25.2 μl ofpyridine is added, then the reaction medium is cooled down to 0-5° C.and 9.5 μl of diphosgene is added.

The temperature is allowed to rise to 20° C., followed by diluting withdichloromethane, washing the reaction medium with a 10% aqueous solutionof tartaric acid then with water.

The organic phase is decanted and dried over sodium sulphate. Then thesolvent is evaporated off under reduced pressure.

In this way 43.8 mg of crude product is obtained which is purified bychromatography on silica eluting with a dichloromethane/ethyl acetatemixture 90/10.

34.9 mg of trans phenylmethyl4-[(benzoylamino)oxy]-1-(chlorocarbonyl)-2-pyrrolidinecarboxylate, ofmolecular formula C₂₀H₁₉ClN₂O₅ (M=402.83 g) is recovered.

The corresponding yield is 69%.

13 mg (0.032 mmole) of the compound obtained previously is dissolved in4 ml of toluene.

9 μl of TEA and 7.8 mg of DMAP are added.

The reaction medium is heated to 100° C. overnight.

The solvent is evaporated off under reduced pressure then the residue ispurified by chromatography eluting with dichloromethane.

In this way 4.3 mg of the expected compound, of molecular formulaC₂₀H₁₈N₂O₅ (M=336.37 g) is recovered. The corresponding yield is 40%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.97 (ddd) and 2.85 (ddd): N—O—CH—CH ₂—CH; 3.80 (dd) and 4.14 (dd):N—O—CH—CH ₂—N; 4.75 (dd): N—CH—CH₂; 4.93 (t): N—O—CH—CH₂; 5.04 and 5.31[AB]: O—CH ₂—C₆H₅; 7.77: and 7.25 to 7.50 (m) CH₂—C₆ H ₅ and OC—C₆ H ₅.

IR (CHCl₃): 1735; 1612, 1575, 1496 cm⁻¹

Example 24 3-benzoyl-1,3-diazabicyclo[2.2.2]octan-2-one

2.4 g (10 mmoles) of N-(4-piperidinyl)-benzamide hydrochloride(described in J. Med. Chem. EN. 17 (1974), 736-739), of molecularformula C₁₂H₁₆N₂O is dissolved under a nitrogen atmosphere in 30 ml ofdichloromethane.

The reaction medium is cooled down to 0° C., 2.8 ml of TEA and 0.66 mlof diphosgene are added under agitation.

After a few minutes, dilution is carried out with dichloromethane,followed by washing with a 10% aqueous solution of tartaric acid, thenwith water, decanting the organic phase, drying over magnesium sulphateand the solvent is evaporated off under reduced pressure. Purificationis carried out on silica eluting with a dichloromethane/ethyl acetatemixture 90/10.

1.62 g of the chloride of 4-(benzoylamino)-1-piperidinecarboxylic acid,of molecular formula C₁₃H₁₅ClN₂O₅ (M=266.5 g) is obtained. Thecorresponding yield is 61%.

1.21 g (48 mmoles) of the compound obtained previously is dissolvedunder a nitrogen atmosphere in 37 ml of tetrahydrofuran.

The solution is cooled down to −78° C., then 5 ml of a 1M solution oflithium bis(trimethylsilyl)amide in tetrahydrofuran is added dropwise.

The reaction medium is maintained at −78° C. for 15 minutes, thetemperature is allowed to rise to ambient temperature and the reactionmedium is left to react again for one hour.

The solution is cooled down to 0° C., 720 μl of acetic acid is added. Aprecipitate forms, followed by diluting with ethyl acetate then washingwith a 10% aqueous solution of tartaric acid and with a solution ofphosphate buffer at pH=7.0.

The organic phase is decanted and dried over magnesium sulphate,followed by filtering, then the solvent is evaporated off under reducedpressure. The crude product is purified by chromatography on silicaeluting with a dichloromethane and ethyl acetate mixture 90/10.

In this way 0.214 g of expected compound, of formula C₁₈H₁₄N₂O₂ (M=230g) is obtained crystallized from ethyl ether.

The corresponding yield is 20%.

1H NMR

In the DMSO, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.71 to 2.02 (m): (CH ₂)₂—CHN; 3.14 (t): N—(CH₂)₂; 4.84 (m): (CH₂)₂—CHN;7.39 to 7.65 (m): C₆ H ₅.

IR (CHCl₃): 1735, 1682; 1618, 1602, 1582; 1488 cm⁻¹

MS (positive electrospray) m/z: [2M+Na]⁺=483; [M+Na+CH₃CN]⁺=294;[M+Na]⁺=253

Example 25 trans diphenylmethyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-2-carboxylate

15 ml of dichloromethane and 197 mg (0.633 mmole) of transdiphenylmethyl 5-hydroxy-2-piperidinecarboxylate (described in Rec.Trav. Chim. (1959), 78, 648-658), of molecular formula C₁₉H₂₁NO₃ aremixed together under an inert atmosphere.

The reaction medium is cooled down to 0° C., then 42 μl of diphosgene,177 μl of TEA then 77 mg of DMAP are added successively. The reactionmedium is left to react for 4 hours at ambient temperature.

Then the reaction mixture is washed with a 10% aqueous solution oftartaric acid, then with a saturated aqueous solution of sodiumchloride.

The organic phases are combined and dried over magnesium sulphate, thesolvent is evaporated off under reduced pressure and in this way 195 mgof crude product is obtained which is purified by chromatography onsilica, eluting with dichloromethane containing 0.1% water.

An oil is recovered which crystallizes from a pentane/ethyl ethermixture.

In this way 108 mg of expected compound is recovered in the form ofwhite crystals corresponding to the molecular formula C₂₀H₁₉NO₄(M=337.338 g).

The corresponding yield is 51%.

1H NMR

In CDCl3, at 400 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.86 (m) and 2.03 (m): N—CH—CH₂—CH ₂—CO; 2.27 (m): N—CH—CH ₂—CH₂—CO;3.07 (d) and 3.29 (m): N—CH ₂—CHO; 4.31 (dd): N—CH—CH₂; 4.73 (m): N—CH₂—CHO; 6.93 (s): CO₂—CH—(C₆H₅)₂; 7.27 to 7.41 (m): CH(C₆ H ₅)₂;

IR (CHCl₃): 1788, 1736; 1496 cm⁻¹;

MS (SIMS) m/z: [M+Na]⁺=360, [M+li]⁺=344; [M]⁺=337, 167

Example 26a trans(4-nitrophenyl)methyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-2-carboxylate

66 ml of dichloromethane and 1 g (3.56 mmole) oftrans(4-nitrophenyl)methyl 5-hydroxy-2-piperidine carboxylate ofmolecular formula C₁₃H₁₆N₂O₅ (M=280.282 g) are mixed together under aninert atmosphere.

The reaction medium is cooled down to 0° C., and 0.24 ml of diphosgeneis added. The reaction medium is left to react whilst agitating for 10minutes at 0° C., then left to heat up to ambient temperature. Thesolvent is evaporated off under reduced pressure.

The residue is dissolved in 66 ml of toluene and 0.99 ml of TEA isadded.

The flask is immersed in an oil bath at 110° C. and maintained there for15 minutes, followed by leaving to return to ambient temperature,washing with a 10% aqueous solution of tartaric acid, then with ansaturated aqueous solution of sodium chloride.

The organic phase is dried over magnesium sulphate then the solvent isevaporated off under reduced pressure.

In this way 0.885 g of crude product is obtained which is purified bychromatography on silica eluting with a toluene/ethyl acetate mixture85/15.

In this way 0.184 g of expected compound, of molecular formulaC₁₄H₁₄N₂O₆ (M=306.276 g) is recovered in the form of a yellow oil.

The corresponding yield is 17%.

1H NMR

In CDCl3, at 400 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.92 (m) and 2.07 (m): N—CH—CH₂—CH ₂—CO; 2.22 (m) and 2.30 (m): N—CH—CH₂—CH₂—CO; 3.17 (d) and 3.35 (dm): N—CH ₂—CHO; 4.28 (dd): N—CH—CH₂; 4.79(m): N—CH₂—CHO; 5.33 [AB]: CO₂—CH ₂—C₆H₄NO₂; 7.56 and 8.25 [AA′BB′]:CH₂—C₆ H ₄—NO₂

IR (CHCl₃): 1791, 1745; 1609, 1526, 1495 cm⁻¹;

MS (EI) m/z: [M]⁺=306, 262, 136, 126, 82, 55

Example 26b trans-7-oxo-6-oxa-1-azabicyclo[3.2.1.]octane-2 carboxylicacid

140 mg (0.457 mmole) of the ester obtained in Example 26a, 7 ml ofacetone and 28 mg of Pd/C catalyst at 20% by weight are mixed together.

The reaction medium is left to react whilst agitating for 25 minutesunder a hydrogen atmosphere at normal pressure.

The catalyst is filtered and then the solvent is evaporated off underreduced pressure.

In this way 137 mg of expected compound, of molecular formula C₇H₉NO₄(M=171.152 g) is obtained, in the form of an oil, in mixture with onemole of β-toluidine.

The corresponding yield is 97%.

1H NMR

In DMSO, at 400 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.84 (m) and 1.95 to 2.05 (m): N—CH—CH ₂—CH ₂—CO; 3.13 (d) and 3.24(dd): N—CH ₂—CHO; 4.02 (dd): N—CH—CH2; 4.81 (dm): N—CH2-CHO.

Example 26c trans methyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-2-carboxylate

17.25 mg (0.1 mmole) of the acid obtained in Example 26b is dissolved in3 ml of dichloromethane.

The reaction medium is treated with an excess of diazomethane insolution in dichloromethane, then the solvent is evaporated off underreduced pressure.

In this way 30 mg of crude product is obtained which is purified bychromatography on silica, eluting with a toluene/ethyl acetate mixture90/10.

6.7 mg of expected compound (M=485.187 g) is recovered.

The corresponding yield is 36%.

Example 27 cis(4-nitrophenyl)methyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-2-carboxylate

0.802 g (2.034 mmoles) of the trifluoroacetate ofcis(4-nitrophenyl)methyl 5-hydroxy-2-piperidine-carboxylate (describedin Rec. Trav. Chim. (1959), 78, 648-658), of molecular formulaC₁₃H₁₆N₂O₅,CF₃CO₂H (M=394.303 g) is introduced under a nitrogenatmosphere, into 40 ml of dichloromethane and the reaction medium iscooled down to 0° C. 0.135 ml of diphosgene is added. Agitation iscarried out for 15 minutes at 0° C., the temperature is left to rise toambient temperature and agitation is continued for 35 minutes.

The solvent is evaporated off under reduced pressure.

This product is dissolved in 40 ml of toluene and 1.1 ml oftriethylamine. The reaction mixture is taken to 100° C. for 35 minutes,then left to cool down to ambient temperature, followed by washing withwater then with a solution of phosphate buffer at pH=7.

The organic phase is dried over sodium sulphate and the solvent isevaporated off under reduced pressure.

In this way 0.56 g of a crude product is obtained which is purified bychromatography on silica, eluting with a dichloromethane/acetone mixture95/5.

In this way 110 mg of the expected compound, of molecular formulaC₁₄H₁₄N₂O₆, (M=306.275 g) is recovered, in the form of an oil.

The corresponding yield is 17%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.80 to 1.94 and 2.10 to 2.45: N—CH—CH ₂—CH ₂—CO; 3.07 (d), 3.04 (dm)and 3.86 (dd): CH—N—CH ₂; 4.80 (t): O═C—O—CH; 5.28 and 5.43 [AB]:O═C—O—CH ₂—C₆H₅; 7.61 and 8.24 [AA′BB′]C₆ H ₄NO₂

IR (CHCl₃): 1801, 1794, 1745, 1704; 1609, 1525, 1498 cm⁻¹.

MS (EI) m/z: [M]⁺=306, 262, 136, 126, 83, 55

Example 28a 1-propenyltriphenylphosphonium salt of trans phenylmethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate Stage Acis phenylmethyl 5-hydroxy-1-(trifluoroacetyl-2-piperidinecarboxylate

6.19 g (22.77 mmoles) of the hydrochloride of phenylmethyl5-hydroxy-2-piperidinecarboxylate, of molecular formula C₁₃H₁₈ClNO₃(M=271.746 g) (described in Rec. Trav. Chim. (1959), 78, 648-658) isdissolved under an inert atmosphere in 80 ml of anhydrousdichloromethane.

The reaction medium is cooled down to 5° C. and 9.5 ml of TEA is addedthen, 6.46 ml of trifluoroacetic anhydride is added dropwise.

The reaction medium is left to react under agitation at 5° C. for onehour, then diluted with dichloromethane, followed by washingsuccessively with a 10% solution of tartaric acid, an aqueous solutionof phosphate buffer at pH=7 and an aqueous solution of sodium chloride.

The organic phase is decanted and dried over magnesium sulphate. Thenthe solvent is evaporated of f under reduced pressure.

In this way 10 g of a red oil is obtained which is dissolved in 100 mlof methanol. The reaction medium is cooled down to about 10° C., and 6.8g (78 mmoles) of sodium hydrogen carbonate in solution in 100 ml ofwater is added slowly, at 20° C. maximum.

The reaction medium is left to react under agitation at 20° C. for 30minutes, then extracted with dichloromethane.

The organic phase is decanted, washed with an saturated aqueous solutionof sodium chloride and dried over magnesium sulphate.

The solvent is evaporated off under reduced pressure and in this way 7.6g of an oil orange oil is collected which is purified by chromatographyon silica, eluting with a dichloromethane/ethyl acetate mixture 95/5.

In this way 6 g of expected compound of molecular formula C₁₅H₁₆F₃NO₄(M=331.294 g) is recovered. The corresponding yield is 68%.

Stage B transphenylmethyl-5-[(2-propenyloxy)amino]-1-(trifluoroacetyl)-2-piperidinecarboxylate

1.74 g (5.26 mmoles) of the alcohol obtained previously is introducedinto 29 ml of acetonitrile. The reaction medium is cooled down to −40°C. and 0.61 ml of 2,6-lutidine (C₅H₃N(CH₃)₂) then 0.91 ml of trifluoromethanesulphonic anhydride is added at this temperature.

The reaction medium is left to react under agitation for 30 minutes at−40° C. 0.7 ml (10.52 mmoles) of O-allyl-hydroxylamine is then added,still at −40° C., over one minute.

The reaction medium is left to return to 0° C. then 0.61 ml of 2.6lutidine is added and the medium is left to react overnight (15 hours),at approximately 5° C., then again for 2 hours at 20° C., followed bydiluting with dichloromethane, washing with an aqueous solution ofsodium hydrogen carbonate, then with a 10% aqueous solution of tartaricacid and with a saturated aqueous solution of sodium chloride.

The organic phase is decanted, dried over magnesium sulphate and thesolvent is evaporated off under reduced pressure.

In this way 2.1 g of a yellow oil is obtained which is purified bychromatography on silica, eluting with a toluene/ethyl acetate mixture90/10.

1.23 g of expected compound of molecular formula C₁₈H₂₁F₃N₂O₄ (M=386.374g) is recovered.

The corresponding yield is 61%.

Stage C trans phenylmethyl5-[(2-propenyloxy)amino]-2-piperidinecarboxylate

1.41 g (3.65 mmoles) of the compound obtained previously is dissolvedunder an inert atmosphere in 25 of anhydrous methanol.

The reaction medium is cooled down to 0-5° C., then 3 additions of 145mg of NaBH₄ are carried out spaced 45 minutes apart.

The reaction medium is then acidified to pH=2 with a 1N aqueous solutionof hydrochloric acid cooled down to 5° C. beforehand.

Extraction is carried out with ethyl acetate.

The aqueous phase is cooled down to 50° C., 100 ml ethyl acetate isadded, followed by treating with a saturated solution of sodiumcarbonate until a pH of 8.5 to 9 is obtained and extracting the aminewith ethyl acetate. The organic phase is washed with a saturated aqueoussolution of sodium chloride, then dried over of magnesium sulphate andconcentrated by evaporation of the solvent under reduced pressure.

In this way 0.628 g of expected product of molecular formula C₁₆H₂₂N₂O₃(M=290.364 g) is obtained.

The corresponding yield is 59%.

Stage D trans phenylmethyl7-oxo-6-(2-propenyloxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

103 mg (0.35 mmoles) of the amine obtained previously is dissolved underan inert atmosphere in 35 ml of anhydrous dichloromethane.

The solution is cooled down to about 0-5° C., and 0.1 ml of TEA, then 21μl of diphosgene are added dropwise, at this temperature.

The reaction medium is left to react under agitation for 15 minutes at0-5° C., then the temperature is allowed to rise to 20° C., and 42 mg ofDMAP is added. Agitation is continued at 20° C. for approximately 5hours, followed by diluting with dichloromethane, washing with a 10%aqueous solution of tartaric acid, then with water.

The organic phase is dried over magnesium sulphate and concentrated byevaporation of the solvent under reduced pressure.

In this way 70 mg of crude product is obtained which is purified bychromatography on 5 g of silica, eluting with a dichloromethane/methanolmixture 98/2.

48 mg of expected product of formula C₁₇H₂₀N₂O₄ (M=316.36 g) isrecovered.

The corresponding yield is 43%.

IR (CHCl₃): 1750; 1642; 1600, 1496 cm⁻¹.

MS (positive electrospray) m/z: [M+Na+CH3CN]⁺=380; [M+Na]⁺=339;[M+H]⁺=317.

Stage E 1-propenyltriphenylphosphonium salt of trans phenylmethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

202 mg (0.638 mmoles) of the compound obtained in Stage D is dissolvedunder an inert atmosphere in 5.5 ml of anhydrous dichloromethane.

73 μl of acetic acid, 369 mg of Pd(P(C₆H₅)₃)₄A are then added at 20° C.to the solution obtained.

After agitation for 30 minutes at ambient temperature, theN-hydroxy-urea formed is treated with 5.5 ml of pyridine and 358 mg ofSO₃-pyridine complex.

The reaction medium is left to react under agitation for 18 hours at 20°C., followed by concentrating by evaporation of the solvent underreduced pressure, taking up in 50 ml of dichloromethane and washing withwater. The organic phase is dried over magnesium sulphate and thedichloromethane is evaporated off under reduced pressure.

In this way 650 mg of crude product is obtained which is purified bychromatography on silica, eluting with a dichloromethane/acetone mixture60/40 containing 0.1% by volume of TEA.

In this way 280 mg of the phosphonium salt of the expected compound, ofmolecular formula C₃₅H₃₅N₂O₇PS (M=646.705 g) is recovered.

The corresponding yield is 68%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

2.05 (m), 2.22 (dm) and 2.33 (m): N—CH—CH ₂—CH ₂; 2.95 (d) and 3.30(dt); O═C—N—CH ₂; 4.10 (m) and 4.32 (m): O═C—N—CH and O═C—N—CH₂—CH; 5.12(s): COO—CH ₂—C₆H₅; 7.36: C₆ H ₅ and 2.30 (m): CH ₃—CH═CH; 6.65 and 7.20CH₃—CH═CH; 7.65-7.85 P(C₆ H ₅)₃.

IR (CHCl₃): 1746; 1638, 1605, 1587, 1495 cm⁻¹.

MS (negative and positive electrospray) m/z: [Manion]⁻=355;[Mcation]⁺=303.

Example 28b Sodium salt of trans phenylmethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

236 mg (0.364 mmoles) of the phosphonium salt obtained in Stage E ofExample 28a is dissolved in 0.8 ml of tetrahydrofuran and 4 drops ofwater.

The solution obtained is passed through a column of DOWEX 50WX8 resin inNa+ form, eluting with water.

After lyophilization, 127 mg of the expected sodium salt, of molecularformula C₁₄H₁₅N₂O₇SNa (M=378.339 g). is obtained

The corresponding yield is 92%.

1H NMR

In DMSO, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.65 to 2.02: N—CH—CH ₂—CH ₂; 2.91 (d) and 3.04 (dt): O═C—N—CH ₂; 4.00to 4.05: O═C—N—CH and O═C—N—CH₂—CH; 5.20 [AB]: COO—CH ₂—C₆H₅; 7.39 (m):C₆ H ₅.

IR (Nujol): 1744; 1495 cm⁻¹.

MS (negative electrospray)_m/z; [M]⁻=355.

Example 28c trans phenylmethyl7-oxo-6-[(phenylsulphonyl)oxy]-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

48 mg (0.152 mmoles) of the derivative obtained in Stage D of Example28a is dissolved in 1.2 ml of dichloromethane.

26 μl of acetic acid then 88 mg of Pd (PPh₃)₄ are added to it at 20° C.,and the reaction medium is left to react for 2 hours at 200 underagitation, followed by diluting by the addition of toluene and thesolvents are evaporated off under reduced pressure.

1.5 ml of dichloromethane, 25 μl of pyridine and 24 μl ofbenzenesulphonyl chloride are added to the crude product obtained.

The reaction medium is left to react at 20° C. under agitation for 1hour then 12.5 μl of pyridine and 10 μl of benzenesulphonyl chloride areadded.

Agitation is carried out for 15 minutes at 20° C., followed by dilutingwith dichloromethane, washing successively with a 10% aqueous solutionof tartaric acid, a solution of phosphate buffer at pH=7 and with asaturated aqueous solution of sodium chloride.

The aqueous phase is dried over magnesium sulphate, and the solvent isevaporated off under reduced pressure. 180 mg of a yellow oil isobtained which is purified by chromatography on silica, eluting with adichloromethane/methyl and t-butyl ether mixture 95/5.

In this way 20 mg of expected compound, of molecular formula C₂₀H₂₀N₂O₆S(M=416.456 g) is recovered. The corresponding yield is 31%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.83 (m) and 2.00 to 2.25 (m): N—CH—CH ₂—CH ₂; 3.02 (d) and 3.16 (dm):O═C—N—CH ₂; 4.04 (m) and 4.11 (dd): O═C—N—CH and O═C—N—CH₂—CH; 5.21 (s):COO—CH ₂—C₆H₅; 7.34 (m): C₆ H ₅; 7.56 (m), 7.70 (m) and 8.03 (m): O₂S—C₆H ₅.

IR (CHCl₃): 1780, 1738; 1600, 1585, 1498; 1386, 1193 cm⁻¹.

MS (positive electrospray) m/z: [2M+Na]⁺=855; [M+Na+CH₃CH]⁺=480;[M+Na]⁺=439; [MH]⁺=417.

Example 28d trans pheylmethyl7-oxo-6-[(2-thienylsulphonyl)oxy]-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

Starting from 100 mg (0.316 mmoles) of the compound obtained in Stage Dof Example 28a, the operation is carried out in a similar manner to thatwhich has just been described, except that instead of usingbenzenesulphonyl chloride, 2 thienyl sulphonyl chloride is used.

In this way 8 mg of expected compound, of molecular formula C₁₈H₁₈N₂O₆S₂(M=422.481 g) is recovered. The corresponding yield is 30%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.84 (m) and 2.10 to 2.25: N—CH—CH ₂—CH ₂; 3.02 (d) and 3.24 (dt):O═C—N—CH ₂; 4.06 (m): O═C—N—CH₂—CH; 4.14 (dd): O═C—N—CH; 5.22 (s):COO—CH ₂—C₆H₅; 7.17 (dd): SO₃—C—S—CH═CH; 7.35 (bs): C₆ H ₅; 7.80 (dd):SO₃—C═CH; 7.87 (m): SO₃—C—S—CH.

IR (CHCl₃): 1780, 1739; 1600, 1503, 1495 cm⁻¹.

MS (positive electrospray) m/z: [M+Na+CH₃CN]⁺=867; [2M+Na]⁺=445; 339,298, 91.

Example 28e trans phenylmethyl6-(2-hydroxy-2-oxoethoxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylateStage A trans phenylmethyl7-oxo-6-[2-oxo-2-(2-propenyloxy)ethoxy]-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

48 mg (0.15 mmoles) of the compound obtained in Stage D of Example 28ais dissolved under an inert atmosphere in 1.5 ml of anhydrousdichloromethane.

18 μl of acetic acid then 88 mg of Pd(P(C₆H₅)₃)₄ are added at 20° C. andthe reaction medium is left under agitation for 1 hour at 20° C.,followed by filtering on silica, eluting with a dichloromethane/t-butyland methyl ether of mixture 7/3.

The solvent is evaporated off under reduced pressure and 70 mg ofhydroxy urea is obtained which is taken up in 2 ml of dichloromethane,then 85 μl of TEA and 64 μl of allyl bromoacetate are added.

Agitation is carried out to 20° C. for 3 hours and 30 minutes, followedby washing successively with a 10% aqueous solution of tartaric acid,with an aqueous solution of phosphate buffer at pH=7 and with water.

The organic phase is dried and the solvent is evaporated off underreduced pressure.

In this way 60 mg of crude product is obtained which is chromatographedon silica eluting with a dichloromethane/t-butyl and methyl ethermixture 90/10 containing 0.1% TEA.

22 mg of molecular formula C₁₉H₂₂N₂O₆, (M=374.396 g) is recovered. Thecorresponding yield is 39%.

Stage B trans phenylmethyl6-(2-hydroxy-2-oxoethoxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

22 mg (0.0587 mmoles) of the compound obtained previously is dissolvedunder an inert atmosphere in 1 ml of anhydrous dichloromethane.

10 μl of acetic acid and 34 mg of Pd(P(C₆H₅)₃)₄ are added at 20° C. andthe reaction medium is left to react under agitation at 20° C. for 30minutes.

The reaction medium is concentrated and taken up in toluene in order toeliminate the acetic acid.

In this way 49 mg of crude product is obtained to which 2 ml ofphosphate buffer of pH 7 is added, then it is washed twice with 1 ml ofdichloromethane.

The solvent is evaporated off and 46 mg of crude product is obtainedwhich is purified by chromatography on silica, eluting firstly with adichloromethane/t-butyl and methyl ether mixture 90/10 then with adichloromethane/ethanol mixture 60/40.

In this way 4.5 mg of expected compound, of molecular formulaC₃₇H₃₇N₂O₆P (M=636.691 g) is obtained. The corresponding yield is 12%.

Example 29a trans(4-nitrophenyl)methyl6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate Stage A cis1-(1,1-dimethylethyl) and2-[(4-nitrophenyl)methyl]5-(methylsulphonyl)oxy-1,2-piperidinedicarboxylate

11.25 g (29.5 mmoles) of cis 1-(1,1-dimethylethyl) and2-[(4-nitrophenyl)methyl]5-hydroxy-1,2-piperidinedicarboxylate(described in Rec. Trav. Chim. (1959), 78, 648-658), of molecularformula C₁₈H₂₄N₂O₇ (M=380.398 g) is dissolved under an inert atmospherein 112 ml of dichloromethane.

The reaction medium is cooled down to 0-5° C., then 5 ml of TEA then2.44 ml of methanesulphonyl chloride are introduced successively.

The temperature is allowed to return to 20° C. under agitation and themedium is left to react for 1 hour, followed by diluting withdichloromethane, washing twice with water, drying over sodium sulphate,and the solvent is evaporated off under reduced pressure.

In this way 16 g of a crude oil is obtained which is purified bychromatography on silica, eluting with dichloromethane containing 2%ethyl acetate.

9.14 g of expected product of molecular formula C₁₉H₂₆N₂O₉S (M=458.491g) is recovered. The corresponding yield is 67%.

Stage B trans 1-(1,1-dimethylethyl) and2-[(4-nitrophenyl)methyl]5-azido-1,2-piperidinedicarboxylate

11.1 g (24.2 mmoles) of the mesylate obtained previously is dissolvedunder an inert atmosphere in 111 ml of dimethylformamide.

Then 1.73 g of sodium nitride NaN₃ is added.

The reaction medium is heated under agitation at 80° C. and ismaintained at this temperature for 18 hours, then left to return to 20°C. The dimethylformamide is evaporated off under reduced pressure untila small volume is obtained, then dilution is carried out with ethylacetate, followed by washing with a 2N solution of soda, then withwater, drying over magnesium sulphate, then the solvents are evaporatedoff under reduced pressure.

The crude oil obtained is purified by chromatography on silica elutingwith dichloromethane containing 2% ethyl acetate.

In this way 7.34 g of expected compound, of molecular formula C₁₈H₂₃N₅O₆(M=405:413 g) is obtained in the form of a yellow oil whichcrystallizes.

The corresponding yield is 75%.

Stage C trans 1-(1,1-dimethyl-ethyl) and2-[(4-nitrophenyl)methyl]5-amino-1,2-piperidinedicarboxylate

7.34 g (18.1 mmoles) of the azide obtained previously is introduced into150 ml of tetrahydrofuran and 30 ml of water.

7.2 g of triphenylphosphine is added, then the reaction medium is leftto react overnight under agitation at 20° C.

Then the solvent is evaporated off under reduced pressure and twoentrainments are carried out with ethyl acetate.

In this way a dry extract is obtained which is purified bychromatography on silica, eluting with dichloromethane containing 5%methanol.

5.62 g of expected compound, of molecular formula C₁₈H₂₅N₃O₆ (M=379.416g) is recovered. The corresponding yield is 82%.

Stage D trans 1-(1,1-dimethylethyl) and2-[(4-nitrophenyl)methyl]5-(benzoylamino)-1,2-piperidinedicarboxylate

700 mg (1.84 mmole) of the amine obtained previously is dissolved in 8ml of dichloromethane.

The reaction medium is cooled down to 0° C., then 257 μl of TEA then 214μl of benzoyl chloride are introduced.

The temperature is allowed to return to 20° C.

After reaction for 40 minutes, dilution is carried out withdichloromethane, followed by washing with a saturated solution of sodiumhydrogen carbonate, then with water, drying over sodium sulphate and thesolvent is evaporated off under reduced pressure.

In this way 867 mg of expected compound, of molecular formula C₂₅H₂₉N₃O₇(M=483.525 g) is obtained. The corresponding yield is 97%.

Stage E hydrochloride of trans(4-nitrophenyl)methyl5-(benzoylamino)-2-piperidine carboxylate

861 mg (8 mmole) of the amide obtained previously, 9 ml of methanol, and2.3 ml of a solution of gaseous hydrogen chloride at 8 mol/l in methanolare mixed together.

The temperature is allowed to return to 20° C. and the reaction mediumis left to react for 3 hours. Then 1.15 ml of a solution of hydrogenchloride in methanol is added.

Agitation is carried out for 20 minutes at 20° C., then the solvent isevaporated off under reduced pressure.

Then two entrainments are carried out with dichloromethane, then twoentrainments with ethyl ether.

The product crystallizes from ethyl ether.

In this way 715 mg of expected compound of molecular formulaC₂₀H₂₂ClN₃O₅ (M=419.967 g) is obtained.

The corresponding yield is 96%.

Stage F trans(4-nitrophenyl)methyl5-(benzoylamino)-1-(chlorocarbonyl)-2-piperidine carboxylate

1.08 g (2.58 mmole) of the hydrochloride obtained as previously and 11ml of dichloromethane are mixed together.

The suspension obtained is cooled down to about 0-5° C. and 791 μl ofTEA is added, then 161 μl of diphosgene is added to the solutionobtained.

Agitation is carried out for 5 minutes at 0-5° C., then the reactionmedium is left to return to 20° C., and left under agitation for another30 minutes, followed by diluting with dichloromethane, washing with a10% aqueous solution of tartaric acid, then with water, drying oversodium sulphate and the solvent is evaporated off under reducedpressure.

The crude product is purified by chromatography on silica eluting withdichloromethane containing 5% acetone.

969 mg of expected compound of molecular formula C₂₁H₂₀ClN₃O₆ (M=445.862g) is recovered.

The corresponding yield is 84%.

Stage G trans(4-nitro-phenyl)methyl6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

928 mg (2.08 mmoles) of the compound obtained previously and 27 ml oftetrahydrofuran are mixed together under an inert gas.

The solution obtained is cooled down to −78° C. under agitation, then2.1 ml of a 1M solution of lithium bis(trimethylsilyl)amide intetrahydrofuran is introduced.

The reaction medium is left under agitation for 10 minutes at −78° C.then 130 μl of acetic acid is added and agitation is carried out whilstallowing the temperature to rise to 15° C., followed by diluting withethyl acetate then washing successively with a 10% aqueous solution oftartaric acid, with a solution of phosphate buffer at pH=7 and withwater, drying over magnesium sulphate and the solvent is evaporated offunder reduced pressure.

In this way 1.6 g of a dry extract is obtained which is purified bychromatography on silica eluting with a dichloromethane/acetone mixture98/2.

The product is then crystallized from ethyl ether in order to produce204 mg of expected compound, of molecular formula C₂₁H₁₉N₃O₆ (M=409.441g).

The corresponding yield is 24%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.98 (m), 2.22 (m) and 2.40 (m): N—CH—CH ₂—CH ₂; 3.08 (d) and 3.42 (dt):O═C—N—CH ₂; 4.23 (dd): O═C—N—CH; 4.53 (m): O═C—N—CH₂—CH; 5.34 [AB]:COO—CH ₂—C₆H₅; 7.69 (m): 8.25 (m): 7.44 (m) and 7.56 (m): C₆ H ₅ and C₆H ₄NO₂.

IR (CHCl₃): 1763, 1744, 1676; 1609, 1603, 1583, 1526, 1492 cm⁻¹.

MS (EI) m/z: [M]⁺=409, 304, 273, 201, 105, 77.

Example 29btrans-6-benzoyl-7-oxo-1,6-iazabicyclo[3.2.1]octane-2-carboxylic acid

89 mg of the ester obtained in Example 29a, 4 ml of acetone and 6 mg of10% Pd/C catalyst are mixed together.

The reaction medium is left to react under agitation, at 20° C. andunder a hydrogen atmosphere for 2 hours 45 minutes, then the catalyst isfiltered and the filtrate evaporated under reduced pressure.

In this way 88 mg of a resin is obtained which is crystallized from 0.5ml of ethyl ether.

In this way 54 mg of expected compound, of molecular formula C₁₄H₁₄N₂O₄(M=274.278 g) is obtained. The corresponding yield is 91%.

1H NMR

In CDCl3, at 250 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.96 (m), 2.10 (m) and 2.37 (m): N—CH—CH ₂—CH ₂; 3.13 (d) and 3.41 (dm):O═C—N—CH₂; 4.10 (bd): O═C—N—CH; 4.52 (m): O═C—N—CH₂—CH; 7.44 (m): 7.56(tt) and 7.69 (dd) C₆ H ₅.

MS (EI) m/z: M⁺=274, 229, 169, 105, 77.

Example 29c trans methyl6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

2 ml of a solution of diazomethane at 12.7 g/l in dichloromethane isadded under agitation to 28 mg (0.102 mmole) the acid obtained inExample 29b.

The solvent is evaporated off under reduced pressure and the residue ispurified by chromatography on silica eluting with adichloromethane/ethyl acetate mixture 98/2.

18.4 mg of expected compound, of molecular formula C₁₅H₁₆N₂O₄ (M=288.305g) is recovered. The corresponding yield is 63%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.90 to 2.42: N—CH—CH ₂—CH ₂; 3.12 (d) and 3.44 (dt): O═C—N—CH ₂; 3.83(s): CH₃; 4.17 (bd): O═C—N—CH; 4.54 (m): O═C—N—CH₂—CH; 7.44 (t), 7.56(t) and 7.69 (d): C₆ H ₅.

MS (EI) m/z: [M]⁺=288, 229, 183, 155, 105, 77.

Example 29dtrans-6-benzoyl-7-oxo-N-(phenylmethyl)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide

30 mg (0.109 mmole) oftrans-6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acidobtained in Example 29b, 0.5 ml of dichloromethane, 23 mg of EDCI and 13μl of benzylamine are mixed together.

The reaction medium is left to react for 30 minutes under agitationfollowed by diluting with dichloromethane, washing with a 10% aqueoussolution of tartaric acid, decanting and drying the organic phase oversodium sulphate.

The solvent is evaporated off under reduced pressure in order to obtaina crude product which is purified by chromatography on silica elutingwith a dichloromethane/acetone mixture 98/2.

In this way 19.5 mg of expected compound, of molecular formulaC₂₁H₂₁N₃O₃ (M=363.419 g) is obtained. The corresponding yield is 49%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.97 (m), 2.34 (m) and 2.59 (m): N—CH—CH ₂—CH ₂; 2.90 (d), 3.33 (m),3.99 (bd) and 4.50 (m): O═C—N—CH, O═C—N—CH₂—CH, O═C—N—CH ₂, CO—NH—CH₂—C₆H₅; 6.94 (bt): NH; 7.24 to 7.58 (m) and 7.68 (m): C₆ H ₅—CO and C₆ H₅—CH₂.

IR (CHCl₃): 3411, 1763, 1680; 1603, 1583, 1519, 1498 cm⁻¹.

Example 29e6-benzoyl-N—[methyl(phenylmethyl)]-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxamide

The operation is carried out in a similar manner to Example 29d startingfrom 50 mg (0.182 mmole) of the acid obtained in Example 29b and 45 μlof N-methyl-benzylamine.

In this way 12 mg of expected compound, of molecular formula C₂₂H₂₃N₃O₃(M=377.45 g) is recovered. The corresponding yield is 17%.

MS (EI) m/z: [M]⁺=377, 272, 105.

Example 29f6-benzoyl-2-(hydroxymethyl)-1,6-diazabicyclo[3.2.1]octan-7-one

100 mg (364 mmole) oftrans-6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acidobtained in Example 29b is dissolved, under an inert atmosphere, in 3 mlof tetrahydrofuran.

The reaction medium is cooled down to −10° C. and 40 μl ofmethylmorpholine, then 38 μl of ethyl chloroformate are added.

The reaction medium is left to react for 15 minutes at −10° C., then thetemperature is allowed to rise to 0° C. and 27 mg of NaBH₄, then,dropwise, 1.5 ml of methanol are added.

The reaction medium is left under agitation at 0° C. for 2 hours thenleft to return to ambient temperature.

3 ml of water is added, the reaction medium is left under agitation for15 minutes, then a few drops of ammonium chloride are added. Extractionis carried out with ethyl acetate, followed by drying over magnesiumsulphate, filtering and the solvent is evaporated off under reducedpressure.

In this way 85 mg of a crude product is obtained which is purified bychromatography on silica, eluting with a dichloromethane/methanolmixture 98/2.

In this way 25 mg of expected compound, of molecular formula C₁₄H₁₆N₂O₃(M=260.3 g) is recovered. The corresponding yield is 26%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.61 (m, 1H), 2.00 (m, 2H) 2.30 (m, 1H): CH—CH₂—CH₂—CH; 2.19: 3.23 (d)and 3.26 (dt): N—CH ₂; 3.60 (m): N—CH—CH₂—OH; 3.70 (m) and 3.77 (dd):CH—CH ₂—O; 4.56 (m): N—CH—CH₂—N.

MS (SIMS) m/z: [M+Na]⁺=283, [M+H]⁺=261, [M]⁺=260, 229, 105.

Example 30 trans(4-nitrophenyl)methyl6-acetyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

1 g (2.63 mmoles) of the product prepared in Stage C of Example 29 isdissolved in 12 ml of dichloromethane. 250 μl of acetic anhydride isadded, the reaction medium is left to react for 10 minutes underagitation, then diluted with dichloromethane and washed with a saturatedaqueous solution of sodium hydrogen carbonate.

The organic phase is dried over sodium sulphate, followed by evaporatingto dryness under reduced pressure in order to obtain 1.2 g of trans1-(1,1-dimethylethyl) and2-[(4-nitrophenyl)methyl]5-(acetylamino)-1,2-piperidinedicarboxylate, ofmolecular formula C₂₀H₂₇N₃O₇ (M=421.453 g).

This product is used without purification in stages similar to Stages Eto G of Example 29 and in this way 14 mg of expected compound, ofmolecular formula C₁₆H₁₇N₃O₆ (M=347.330 g) is collected. Thecorresponding yield is 17%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.87 (m), 2.00 to 2.30 (m): N—CH—CH ₂—CH ₂; 2.54 (s): N—CO—CH ₃; 2.95(d) and 3.21 (m): O═C—N—CH ₂; 4.26 (bd): O═C—N—CH; 4.55 (m):O═C—N—CH₂—CH; 5.34 [AB]: CO₂—CH ₂—C₆H₄; 7.57 and 8.25 [AA′BB′]: C₆ H₄—NO₂.

MS (EI) m/z: [M]⁺=347, 304, 211, 169, 125, 43.

Example 31 trans(4-nitrophenyl)methyl and 2-propenyl7-oxo-1,6-diazabicyclo[3.2.1]octane-2,6-dicarboxylate

1.24 g (3.278 mmoles) of the product prepared in Stage C of Example 29ais dissolved, under a nitrogen atmosphere, in 8 ml of dichloromethane.

The solution is cooled down to 0° C., then 0.45 ml of TEA then 0.35 mlof allyl chloroformate are added dropwise.

The reaction medium is maintained at 0° C. for 15 minutes, then is leftto react under agitation for 1 hour at ambient temperature, followed bydiluting with 20 ml of dichloromethane, washing with an aqueous solutionof sodium bicarbonate, and twice with water, drying over magnesiumsulphate, and the solvent is evaporated off under reduced pressure.

In this way 1.5 g of trans 1-(1,1-dimethylethyl) and2-[(4-nitrophenyl)methyl]5-[[(2-propenyloxy)carbonyl]amino]-1,2-piperidinedicarboxylateof molecular formula C₂₂H₂₈N₃O₈, (M=462.486 g) is obtained.

The corresponding yield is 99%.

This product is used in stages similar to Stages E to G of Example 29aand in this way 30.6 mg of expected compound, of molecular formulaC₁₈H₁₉N₃O₇, (M=389.368 g) is obtained in the form of a white solid. Thecorresponding yield is 40%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.91 (m), 2.00 to 2.29 (m): N—CH—CH ₂—CH ₂; 2.98 (d) and 3.25 (bd):O═C—N—CH ₂; 4.27 (t) O═C—N—CH; 4.37 (bs): O═C—N—CH₂—CH; 4.77 (bd):COO—CH ₂—CH═; 5.33 (s): COO—CH ₂—C₆H₄; 5.29 to 5.46: CH ₂═CH; 5.98 (m):CH₂═CH; 7.96 and 8.29 [AA′BB′]: C₆ H ₄—NO₂.

IR (CHCl₃): 1801, 1775, 1738, 1724; 1649; 1608, 1595, 1526 cm⁻¹.

MS (positive electrospray) m/z: [2M+Na]⁺=801, [M+Na+CH₃CN]⁺=453,[M+Na]⁺=412

Example 31a trans phenylmethyl6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

200 mg of trans phenylmethyl5-(benzoylamino)-1-(chlorocarbonyl)-2-piperidinecarboxylate, ofmolecular formula C₂₁H₂₁ClN₂O₄ (M=400.87 g), prepared in a similarmanner to Stages A to F of Example 29a and 6 ml of anhydroustetrahydrofuran are mixed together under an inert atmosphere and thereaction medium is cooled down to −78° C.

0.55 ml of a 1M solution of lithium bis (trimethylsilyl)amide intetrahydrofuran is added dropwise.

The reaction medium is left to react under agitation at −78° C. for 10minutes then 25 μl of acetic acid is added.

The temperature is allowed to rise to ambient temperature, then thereaction medium is poured into 10 ml of a 10% aqueous solution oftartaric acid, followed by extracting with ethyl acetate, washing withan aqueous solution of phosphate buffer at pH=7, then with water, dryingover magnesium sulphate and bringing to dryness by evaporation of thesolvent under reduced pressure.

In this way 158 mg of a crude product is obtained which is purified bychromatography on silica, eluting with a dichloromethane/acetone mixture98/2.

In this way 70 mg of expected compound, of molecular formula C₂₁H₂₀N₂O₄(M=364.40 g) is recovered. The corresponding yield is 39%.

1H NMR

In CDCl3, at 400 MHz, chemical shifts of the peaks in ppm andmultiplicity:

2.15 (m) and 2.25 (m): NCH—CH₂—CH ₂—CH—CO₂; 1.94 (m) and 2.36 (m):NCH—CH ₂—CH₂—CH—CO₂; 4.20 (d) N—CH—CO₂; 4.50 (q): NCH—CH₂—CH₂—CH—CO₂;3.08 (d) and 3.40 (dt): N—CH ₂; 5.25 [AB]: CO₂—CH ₂—C₆H₅; 7.38 (bs):CH₂—C₆ H ₅; 7.43 (bt) and 7.55 (bt) and 7.69 (bd) C₆ H ₅—CO.

IR (CHCl₃): 1764, 1744, 1675; 1602, 1584, 1498 cm¹.

MS (SIMS) m/z: [M+Na]⁺=387, [M+H]⁺=365, 259, 257, 229, 105, 91.

Example 31b phenylmethyl6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-2-ene-2-carboxylate

46 mg (0.126 mmoles) of the product obtained in Example 31a and 0.5 mlof anhydrous tetrahydrofuran are mixed together, under a nitrogenatmosphere.

The reaction medium is cooled down to −70° C. and 0.31 ml of 1M lithiumbis(trimethylsilyl)amide in tetrahydrofuran is added.

The reaction medium is left to react for 2 hours at −70° C., then thetemperature is allowed to rise to −15° C. and 0.41 ml of a solution ofC₆H₅—SeCl at 0.7 mol/l in THF is added at this temperature.

The reaction medium is left under agitation at −15° C. for 15 minutes,then left to return to ambient temperature over 15 minutes and pouredinto a mixture of water and ice containing a few drops of a saturatedaqueous solution of sodium bicarbonate.

Extraction is carried out with ethyl acetate, followed by washing withwater, drying and the solvent is evaporated off under reduced pressure.

The residue is purified by chromatography on silica eluting with adichloromethane/acetone mixture 98/2 and in this way 15 mg ofphenylmethyl6-benzoyl-7-oxo-2-(phenylselenyl)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,of molecular formula C₂₇H₂₄N₂O₄Se (M=519.46 g) is collected. Thecorresponding yield is 23%.

15 mg (0.029 mmole) of the compound obtained previously and 0.3 ml ofdichloromethane are mixed together.

The reaction medium is cooled down to 0° C. and 15 mg ofmeta-chloroperbenzoic acid in solution in 0.15 ml of dichloromethane isadded.

The reaction medium is left under agitation at 0° C. for 15 minutes,then left to return to ambient temperature, followed by pouring intoapproximately 20 ml of water, extracting with dichloromethane andwashing the organic phase with an aqueous solution of phosphate bufferat pH=7, drying over magnesium sulphate, filtering and the solvent isevaporated off under reduced pressure.

In this way 15 mg of crude product is obtained which is purified onsilica eluting with a dichloromethane/acetone mixture 98/2.

In this way 5 mg of expected compound, of molecular formula C₂₁H₁₈N₂O₄(M=362.39 g) is recovered. The corresponding yield is 48%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity: 2.66 (td) and 2.99 (tdd): N—CH—CH ₂; 3.03 (d) and 3.77(ddd): N—CH ₂; 4.76 (tt): N—CH; 5.23 [AB]: CO2-CH ₂—C₆H₅; 7.02 (dt):N—C═CH; 7.30 to 7.38 (m): CH₂—C₆ H ₅; 7.42 (tm), 7.54 (tm) and 7.62(dm); C₆ H ₅—CO;

Example 31c6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-2-ene-2-carboxylic acid

20 mg (0.055 mmole) of the product obtained in Example 31b is mixed, 0.4ml of acetone and 4 mg of 10% Pd/C catalyst are added.

The reaction medium is placed under a hydrogen atmosphere and left toreact for 3 hours under vigorous agitation.

Filtration is carried out and the catalyst is washed with acetone thenwith methanol. The filtrate is evaporated under reduced pressure.

In this way 14 mg of expected compound, of molecular formula C₁₄H₁₂N₂O₄(M=272.4 g) is obtained. The corresponding yield is 93%.

MS (EI) m/z: [M]⁺: 272, 105.

Example 32a trans 2-propenyl7-oxo-6-(2-phenylmethoxy)-1,6-diaza-bicyclo[3.2.1]octane-2-carboxylateStage A cis 2-propenyl5-hydroxy-1-[(trifluoroacetyl)-2-piperidinecarboxylate

17 g (0.059 mole) of cis 1-(1,1-dimethylethyl) and2-(2-propenyl)5-hydroxy-1,2-piperidinedicarboxylate (described in Rec.Trav. Chim. (1959), 78, 648-658), of molecular formula C₁₄H₂₃NO₅(M=285-3431 g) is dissolved in 17 ml ethyl acetate.

A solution of 51 ml of hydrogen chloride in ethyl acetate at 150 g/l isadded at 0° C.

The reaction medium is left to return to ambient temperature and left toreact under agitation for 1 hour 30 minutes.

The ethyl acetate is evaporated under reduced pressure, followed bytaking up in ethyl ether, which is in turn eliminated under reducedpressure.

In this way 12 g of a pale yellow solid is obtained which is mixed with200 ml of tetrahydrofuran. The reaction medium is cooled down to 0° C.,then 37.6 ml of TEA is added.

The temperature is maintained at 0° C., then 16.8 ml of trifluoroaceticanhydride is added slowly.

The temperature is allowed to rise to 20° C. and the reaction medium isleft to react for another 20 minutes under agitation.

Then 20 ml of water is added.

The solution obtained is agitated for 1 hour at ambient temperature andpoured into 300 ml of water, followed by extracting with ethyl acetate,washing with water, drying over sodium sulphate, and the solvent isevaporated off under reduced pressure.

15.7 g of crude product is obtained which is purified by chromatographyon silica eluting with a dichloromethane/ethyl acetate mixture 90/10.

In this way 12.3 g of expected compound, of molecular formulaC₁₁H₁₄F₃NO₄ (M=281.23 g), is obtained in the form of a yellow oil. Thecorresponding yield is 73%.

Stage B trans 2-propenyl5-[(phenylmethoxy)amino]-1-(trifluoroacetyl)2-piperidinecarboxylate

10.9 g (38.7 mmoles) of the compound obtained in Stage A and 150 ml ofacetonitrile are mixed together.

The pale yellow solution obtained is cooled down to −30° C., then, 4.94ml of 2,6-lutidine and 6.7 ml of trifluoromethanesulphonic anhydride areadded. Agitation is carried out for 15 minutes, then, still at −30° C.,9.57 g of O-benzylhydroxylamine is added.

At end of the addition, the temperature is left to rise to 0° C. and thereaction medium is left to react for 1 hour at this temperature. Then4.9 ml of 2,6-lutidine is added and the reaction medium is left incontact for 3 days at 0° C., followed by pouring into 500 ml of water,extracting with ethyl acetate, washing successively with water, with anaqueous solution of phosphate buffer at pH=7.0, with a saturated aqueoussolution of sodium chloride, then again with water.

After drying over sulphate sodium, the solvent is evaporated off underreduced pressure.

In this way 23 g of crude product is obtained which is dissolved in 150ml of dichloromethane, followed by washing with a 10% aqueous solutionof tartaric acid, drying over sodium sulphate and the solvent isevaporated off under reduced pressure.

In this way 16.1 g of a yellow oil is recovered which is purified bychromatography on silica.

12.1 g of expected compound, of molecular formula C₁₈H₂₁F₃N₂O₄ (M=386.37g) is recovered in crystallized form. The corresponding yield is 72%.

Stage C trans 2-propenyl 5-[(phenylmethoxy)amino]-2-piperidinecarboxylate

80 ml of methanol is cooled down to −10° C., then 4.15 g (37.8 mmoles)of NaBH₄ is added.

A solution of 10.6 g (27.4 mmoles) of the compound obtained previouslyin 80 ml of methanol is added slowly, under agitation, over 30 minutes,to this mixture whilst maintaining the temperature at −10° C.

Then the temperature is allowed to rise to 0° C., then this temperatureis maintained for 3 hours.

The reaction mixture is poured into 450 ml of ice and water and 150 mlethyl acetate, followed by decanting, washing with water, drying theorganic phase over sodium sulphate and then the solvent is evaporatedoff under reduced pressure.

In this way 8.2 g of a yellow oil is obtained which is dissolved in 80ml of tetrahydrofuran, a solution of 2.43 g of oxalic acid in 25 ml ofTHF is added. The oxalate which crystallizes is filtered and washed witha little THF then dried under reduced pressure and dissolved in asaturated solution of sodium bicarbonate. Extraction is carried out withethyl acetate, followed by washing the organic phase with water, dryingover sodium sulphate and the solvent is evaporated off under reducedpressure.

In this way 4.39 g of expected compound, of molecular formula C₁₆H₂₂N₂O₃(M=290.36 g), is obtained in the form of an oil which crystallizes whenthe temperature is below 20° C. The corresponding yield is 55%.

Stage D trans 2-propenyl7-oxo-6-(2-phenylmethoxy)-1,6-diaza-bicyclo[3.2.1]-octane-2-carboxylate

3.2 g (11 mmoles) of the oil obtained previously is dissolved under anitrogen atmosphere in 500 ml of acetonitrile.

The solution obtained is cooled down to 0° C. using an ice bath and 3.37ml of TEA, then 0.796 ml of diphosgene, and 1.48 g of DMAP are added.

The temperature is allowed to rise to 20° C. and the reaction medium isleft to react for 2 hours under agitation.

Then the reaction mixture is poured into 200 ml of a 0.1 N aqueoussolution of hydrochloric acid, 400 ml of water is added, followed byextracting with dichloromethane, washing with water and drying oversodium sulphate.

Then the solvent is evaporated off under reduced pressure so as toobtain 3.1 g of expected compound, of molecular formula C₁₇H₂₀N₂O₄(M=316.36 g), in the form of crystals. The corresponding yield is 89%.

1H NMR

1.66 (m) and 2.00 to 2.16 (m) O═C—CH—CH ₂—CH ₂; 2.94 (d) and 3.07 (dt)N—CH ₂; 3.31 (m) N—CH₂—CH; 4.14 (dd) O═C—CH, 4.68 (dt) CH ₂—CH═CH₂; 4.90and 5.06 [AB] CH ₂—C₆H₅; 5.26 (dq) and 5.34 (dq) CH₂—CH═CH ₂; 5.92 (m)CH₂—CH═CH₂; 7.37 to 7.42 (m) C₆ H ₅.

IR (CHCl₃): 1748; 1646; 1496 cm⁻¹.

MS (positive electrospray) m/z: [2M+Na]⁺=655, [M+Na+CH₃CN]⁺=380,[M+Na]⁺=339, [M+H]⁺=317, 289, 91.

Example 32btrans-7-oxo-6-(phenylmethoxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylicacid and its cyclohexylamine salt

2.21 g (6.98 mmoles) of the compound obtained in Example 32a isdissolved under a nitrogen atmosphere in 44 ml of dichloromethane.

A 0.5M solution of sodium ethyl-hexanoate in ethyl acetate is added.

Then 242 mg of tetrakistriphenylphosphine palladium is added in one go,then the reaction medium is maintained under agitation for 1 hour,followed by diluting with 22 ml ethyl acetate, pouring into 75 ml of asaturated solution of NaH₂PO₄, extracting with ethyl acetate and dryingthe organic phase over sodium sulphate. The solvent is evaporated offunder reduced pressure in order to obtain 3.5 g of a yellow residuewhich is dissolved in a mixture of 11 ml of ethyl acetate and 0.8 ml ofcyclohexylamine.

The crystallized cyclohexylamine salt is separated by filtration andwashed with ethyl ether, then the solvent is evaporated off underreduced pressure. In this way a total 2.51 g of crystallized salt isobtained which is dissolved in 25 ml of a saturated aqueous solution ofNaH₂PO₄. After extraction with ethyl acetate, the organic phases arecombined and dried over sodium sulphate, then the solvent is evaporatedoff under reduced pressure.

In this way 1.82 g of expected compound of molecular formula C₁₄H₁₆N₂O₄(M=276.29 g), is recovered in crystallized form. The corresponding yieldis 94%.

1H NMR

In CDCl3, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.68 (m) and of 2.20 to 2.22 (m): CH—CH ₂—CH ₂—CH; 2.89 (d) and 3.11(ddd): N—CH ₂; 3.34 (dd) N—CH₂—CH, 4.13 (bd): N—CH—C═O; 4.90 and 5.05[AB]: CH ₂—O; 7.32 to 7.43: C₆ H ₅.

MS (SIMS) m/z: [M+Na]⁺=299, [M+H]⁺=277.91.

Example 33a pyridinium salt oftrans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideStage Atrans-7-oxo-6-(phenylmethoxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide

1.1 g (4 mmole) of the compound obtained in Example 32b is dissolved in30 ml of dichloromethane.

0.67 ml of TEA is added to this solution.

The solution is cooled down to 5° C. and 0.57 ml of isobutylchloroformate is added quite quickly.

The reaction medium is maintained under agitation for 20 minutes at 5°C., then 3 ml of concentrated ammonia is added slowly, under vigorousagitation.

Agitation is maintained for one hour at ambient temperature, thereaction medium is diluted with 30 ml of water, followed by extractingwith dichloromethane, washing with water, drying over sodium sulphateand concentrating under reduced pressure.

In this way 1.1 g of expected product of molecular formula C₁₄H₁₇N₃O₃(M=275.31 g) is obtained. The yield is quantitative.

Stage Btrans-6-hydroxy-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxamide

1.1 g of the compound obtained in Stage A, 30 ml of methanol and 300 mgof 10% Pd/C are mixed together.

The reaction medium is placed under a hydrogen atmosphere then themixture is agitated vigorously for 45 minutes.

The catalyst is then filtered, followed by washing with methanol thenwith a dichloromethane/methanol mixture.

The filtrate is evaporated under reduced pressure.

In this way 800 mg of expected product of molecular formula C₇H₁₁N₃O₃(M=185.18 g) is obtained in the form of a colourless foam.

Stage C Pyridinium salt oftrans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide

800 mg of the compound obtained previously and 20 ml of anhydrouspyridine are mixed together under a nitrogen atmosphere.

Then 1.91 g of SO₃-pyridine complex is added.

The mixture is agitated for 20 hours at ambient temperature.

The reaction medium is then filtered and the solvent evaporated offunder reduced pressure.

In this way the expected product of molecular formula C₁₂H₁₆N₄O₆S, C₅H₅N(M=344.35 g) is obtained in the form of a yellow product.

Example 33b Tetrabutylammonium salt oftrans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide

The product obtained previously is introduced into 40 ml of aconcentrated aqueous solution of NaH₂PO₄ so as to obtain a pH of 4.

Extraction is carried out with ethyl acetate then 1.01 g of tetrabutylammonium hydrogen sulphate is added to the aqueous phase

Agitation is carried out for 10 minutes at ambient temperature, followedby extracting with 4 times 300 ml ethyl acetate, drying theorganic-phase over sodium sulphate and concentrating under reducedpressure.

In this way 1.530 g of a colourless foam is obtained which is purifiedby chromatography on silica, eluting with an acetone/dichloromethane/TEAsolvent 50/48/2.

In this way 1.02 g of expected product of molecular formula C₂₃H₄₆N₄O₆S(M=506.71 g), is recovered in the form of a colourless foam. Thecorresponding overall yield is 50%.

Example 33c Sodium salt oftrans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide

The product obtained in Example 33b is dissolved in 7 ml of anacetone/water mixture 1/1 then deposited on a column of 180 g of DOWEX50WX8 resin in Na⁺ form and eluted with water. After evaporation of thewater under reduced pressure, the product crystallizes.

In this way 542 mg of expected compound, of formula C₇H₁₀N₃NaO₆S(M=287.23 g) is obtained. The corresponding yield is 94%.

1H NMR

In DMSO, at 300 MHz, chemical shifts of the peaks in ppm andmultiplicity:

1.55 to 2.10 (3H): CH—CH ₂—CH ₂—CH; 2.91 (d) and 3.02 (bd): N—CH₂; 3.38(bs): N—CH₂—CH; 3.68 (d): N—CH—C═O; 7.23 and 7.44: NH ₂.

MS (negative electrospray) m/z: [M]⁻=264

Examples 34 to 47

The following carboxamides were prepared following an operating methodsimilar to that which is used in Example 33 starting from 110 mg of theacid obtained in Example 32b.

The only difference is that in Stage 1, the reagent used, i.e. theammonia solution, is replaced by a solution of the corresponding amine.

Thus, only the R1 group as defined for formula I varies.

Example 34

Starting from 49 μl of benzylamine, 64 mg of the sodium salt oftrans-7-oxo-N-(phenylmethyl)-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. an overall yield of 38%.

MS (positive electrospray) m/z: [M+Na]⁺=400, [M+H]⁺=378

Example 35

Starting from 43 μl of 2-pyridinemethanamine, 37 mg the sodium salt oftrans-7-oxo-N-(2-pyridinylmethyl)-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. an overall yield of 14%.

MS (positive electrospray) m/z: [M+H]⁺=379

Example 36

Starting from 51.3 mg of 3-pyridineethanamine, 42 mg of the sodium saltoftrans-7-oxo-N-[2-(3-pyridinyl)ethyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. an overall yield of 20%.

MS (positive electrospray) m/z: [M+H]⁺=393

Example 37

Starting from of 51.3 mg of 4-pyridineethanamine, 40 mg of the sodiumsalt oftrans-7-oxo-N-[2-(4-pyridinyl)ethyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 20%.

MS (positive electrospray) m/z: [M+Na]⁺=415, [M+H]⁺=393

Example 38

Starting from 50.2 mg of 2-pyridineethanamine, 45 mg of the sodium saltoftrans-7-oxo-N-[2-(2-pyridinyl)ethyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 23%.

MS (positive electrospray) m/z: [M+H]⁺=393

Example 39

Starting from 58.3 mg of 3-amino-benzamide, 43 mg of the sodium salt oftrans-N-[3-(aminocarbonyl)phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 22%.

MS (negative electrospray) m/z: [M]⁻=383

Example 40

Starting from 58.3 mg of 4-dimethylamino-benzenamine, 65.3 mg of thesodium salt oftrans-N-[4-(dimethylamino)phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 40%.

MS (negative electrospray) m/z: [M]⁻=383

Example 41

Starting from 58.3 mg of 3-dimethylamino-benzenamine, 91 mg of thesodium salt oftrans-N-[3-(dimethylamino)phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 54%.

MS (negative electrospray) m/z: [M]⁻=383

Example 42

Starting from 43 μl of 4-pyridinemethanamine, 24.6 mg of the sodium saltoftrans-7-oxo-N-[(4-pyridinyl)methyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 15%.

MS (negative electrospray) m/z: [M]⁻=355

Example 43

Starting from 44 μl of 3-pyridinemethanamine, 44.7 mg of the sodium saltoftrans-7-oxo-N-(3-pyridinylmethyl)-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 26%.

MS (negative electrospray) m/z: [M]⁻=355

Example 44

Starting from 84 mg (+−)-alpha-amino-benzenepropanamide, 55 mg of thesodium salt oftrans-N-(1-amino-1-oxo-3-phenyl-2-propyl)-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 27%.

MS (negative electrospray) m/z: [M]⁻=411, 321

Example 45

Starting from 46 mg of hydrochloride of 2-amino-acetamide and 61 μl ofTEA, 25 mg of the sodium salt oftrans-N-(2-amino-2-oxoethyl)-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 13%.

MS (negative electrospray) m/z: [M]⁻=321, 249

Example 46

Starting from 64 mg of (3-aminophenyl)-urea, 43 mg of the sodium salt oftrans-N-[3-[(aminocarbonyl)amino]phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 24%.

MS (negative electrospray) m/z: [M]⁻=398, 153, 111

Example 47

Starting from 63 mg of (+−)-alpha-amino-benzeneacetamide, 64 mg of thesodium salt oftrans-N-(2-amino-2-oxo-1-phenylethyl)-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamideis obtained i.e. a yield of 38%

MS (negative electrospray) m/z: [M]⁻=397

Examples 48 to 51

The following compounds were prepared from 110 mg of the compoundobtained in Stage E of Example 32, which is esterified each time withthe appropriate alcohol in order to obtain the final product.

Then, the operation is carried out in a similar manner to that describedin Stages B to E of Example 33.

Example 48

Starting from 31.5 mg of 2-hydroxy-acetamide, 54 mg of the sodium saltof trans 2-amino-2-oxoethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate isobtained i.e. a yield of 32%.

MS (negative electrospray) m/z: [M]⁻=322

Example 49

Starting from 51.7 mg of 4-pyridineethanol, 20 mg of the sodium salt oftrans 2-(4-pyridinyl)ethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate isobtained i.e. a yield of 8.5%.

MS (negative electrospray) m/z: [M]⁻=370

Example 50

Starting from 47.3 mg of 2-pyridineethanol, 47 mg of the sodium salt oftrans 2-(2-pyridinyl)ethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate isobtained i.e. a yield of 23.4%.

MS (negative electrospray) m/z: [M]⁻=370

Example 51

Starting from 57.7 mg of 3-pyridineethanol, 50 mg of the sodium salt oftrans 2-(3-pyridinyl)ethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate isobtained i.e. a yield of 26%.

MS (negative electrospray) m/z: [M]⁻=370

Example 52 Sodium salt of3-methoxy-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one Stage A

10 g (50 mmoles) of 1,1-dimethylethyl 3,5-dioxo-1-piperidinecarboxylateis dissolved in 10 ml of methanol, then 6 g (54 mmoles) ofO-allylhydroxylamineamine hydrochloride is added.

The reaction medium is left under agitation for 3 hours, then thesolvent is evaporated off under reduced pressure.

The residue is taken up in water, followed by extracting withdichloromethane, washing the organic phase with water, then drying itover sodium sulphate.

After filtration and evaporation of the solvent under reduced pressure,10.6 g of 1,1-dimethylethyl5-methoxy-3-[(2-propenyloxy)imino]-3,6-dihydro-1(2H)-pyridinecarboxylateof molecular formula C₁₄H₂₂N₂O₄ (M=282.342 g) is obtained. Thecorresponding yield is 75%.

Stage B

10.6 g (37.6 mmoles) of the product obtained in Stage A and 212 ml ofmethanol are placed in a flask.

The solution is cooled down to −5° C., 37.8 g sodium cyanoborohydide,then 58.2 ml of boron fluoride etherate are added, followed by dilutingwith dichloromethane, pouring onto a mixture of water and 2N soda,extracting with dichloromethane, washing the organic phase with water,drying over sodium sulphate, filtering and the solvent is evaporated offunder reduced pressure. The product obtained is purified bychromatography on silica eluting with an AcOEt/dichloromethane mixture10/90.

In this way 5.5 g of 1,1-dimethylethyl5-methoxy-3-[(2-propenyloxy)amino]-3,6-dihydro-1(2H)-pyridinecarboxylateof molecular formula C₁₄H₂₄N₂O₂ (M=284.36 g) is obtained. Thecorresponding yield is 51%.

Stage C

5.5 g (19.3 mmoles) of the product obtained in Stage B, 27.5 ml ofdichloromethane and 4.2 ml of anisole are introduced into a flask.

Then 27.5 ml of trifluoroacetic acid is added.

The TFA and dichloromethane are eliminated under reduced pressure.

The residue is taken up in water and extraction is carried out 3 timeswith AcOEt. The aqueous phase is rendered basic by the addition ofammonium hydroxide, then extracted with AcOEt.

The organic phases are washed with water, followed by drying over sodiumsulphate, filtering, then the solvent is evaporated off under reducedpressure.

In this way 2.45 g of5-methoxy-N-(2-propenyloxy)-1,2,3,6-tetrahydro-3-pyridinamine ofmolecular formula C₉H₁₆N₂O₂ (M=184.24 g) is obtained. The correspondingyield is 69%.

Stage D

2.45 g (0.0133 mmole) of the product obtained in Stage C is dissolvedunder an inert atmosphere in 826 ml of acetonitrile and the solution iscooled down to 0° C. 0.778 ml of diphosgene is added.

The temperature is allowed to rise to ambient temperature, then 5.56 mlof TEA is added.

Agitation is carried out overnight at ambient temperature, then thesolvent is evaporated off under reduced pressure.

The residue is taken up in water, followed by extracting with AcOEt,washing the organic phase with water, drying over sodium sulphate,filtering, then the solvent is evaporated off under reduced pressure.

The residue is purified by chromatography on silica eluting with anAcOEt/dichloromethane mixture 1/9.

In this way 1.13 g of3-methoxy-6-(2-propenyloxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one ofmolecular formula C₁₀H₁₄N₂O₃ (M=210.23 g) is recovered.

The corresponding yield is 40.3%.

Stage E

In a flask placed under an inert atmosphere, 105 mg (0.5 mmole) of theproduct obtained in Stage D is dissolved in 1.1 ml of dichloromethane,57 μl of acetic acid, then 317 mg of Pd[P(C₆H₅)₃]₄ are added.

After reaction for 1 hour, 1.1 ml of pyridine, then 238 mg ofSO₃-pyridine complex are added.

The reaction medium is left under agitation overnight, then the solventis evaporated off under reduced pressure.

The residue is taken up in water, followed by extracting withdichloromethane, washing with water, drying the organic phase oversodium sulphate, filtering and the solvent is evaporated off underreduced pressure.

The residue is purified by chromatography on silica, eluting with atrichloromethane/acetonitrile mixture 50/50.

In this way 148 mg of the 1-propenyltriphenylphosphonium salt of3-methoxy-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one ofmolecular formula C₂₈H₂₉N₂O₆PS is recovered.

The corresponding yield is 53%.

Stage F

148 mg of the product obtained in Stage E is dissolved in watercontaining 10% THF.

The solution obtained is passed through a column of DOWEX 50WX8 resin inNa⁺ form, eluting with water containing 10% THF.

The product collected is lyophilized in order to obtain 51 mg of theexpected sodium salt, of molecular formula C₇H₉N₂O₆SNa (M=272.21 g).

The corresponding yield is 70%.

1H NMR

3.04 (d) and 3.25 (dd): C═CH—CH—CH ₂—N, 3.41 (d) and 3.71 (dd): N—CH₂—C═CH; 3.47 (s): CH₃—O; 4.20 (dd): C═CH—CH—CH₂—N, 5.19 (bd):C═CH—CH—CH₂—N

MS (negative electrospray) m/z: [M]⁻=249, [M —CH₃]⁻=235

Example 53 Sodium salt of6-(sulphooxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one Stage A

1.03 g (5.2 mmoles) of 1,1-dimethylethyl3,6-dihydro-3-oxo-1(2H)-pyridinecarboxylate of molecular formulaC₁₀H₁₅NO₃ is dissolved in 15 ml of ethanol. 572 mg (5.2 mmoles) ofO-allylhydroxylamineamine, then 1.3 ml of pyridine are added.

The reaction medium is left under agitation for 15 minutes, then 100 mlof dichloromethane is added, followed by washing with a 10% aqueoussolution of tartaric acid, drying the organic phase over magnesiumsulphate, filtering and the solvent is evaporated off under reducedpressure.

In this way 1.36 g of 1,1-dimethylethyl3,6-dihydro-3-[(2-propenyloxy)imino]-1(2H)-pyridinecarboxylate ofmolecular formula C₁₃H₂₀N₂O₃ (M=252.32 g) is obtained.

The corresponding yield is quantitative.

Stage B

The operation is carried out as indicated in Stage A of Example 52starting from 1.38 g of the product obtained in Stage A, 15.1 g ofsodium cyanoborohydide and 8.3 ml of boron trifluoride etherate.

In this way 0.99 g of a mixture of 2/3 of 1,1-dimethylethyl3-[(2-propenyloxy)amino]-1-piperidinecarboxylate and 1/3 of1,1-dimethylethyl 3,6-dihydro-3-[(2-propenyloxy)amino]-1(2H)-pyridinecarboxylate of molecular formula C₁₃H₂₂N₂O₃ (M=254.33 g) is recoveredafter purification.

The corresponding yield is 71%.

Stage C

1.07 g (4.26 mmoles) of a mixture obtained in Stage B is dissolved in 2ml of AcOEt. The reaction medium is cooled down to 0° C., then 5.8 ml ofa 7.3 M solution of hydrogen chloride in AcOEt is added and the mediumis left to react for 2 hours 30 minutes at 0° C.

The solvent is evaporated off under reduced pressure, followed by takingup in ether, filtering the precipitate and drying under reducedpressure.

In this way 560 mg ofN-(2-propenyloxy)-1,2,3,6-tetrahydro-3-pyridinamine dihydrochloride ofmolecular formula C₈H₁₆Cl₂N₂O (M=227.14 g) is obtained.

The corresponding yield is 57%.

Stage D

560 mg (2.46 mmoles) of the product obtained in Stage C is dissolved in6 ml of dichloromethane, then 2.5 ml of 2N soda is added.

After decanting, the aqueous phase is extracted with AcOEt.

The organic phases are combined, followed by drying over magnesiumsulphate and filtering, then the solvent is evaporated off under reducedpressure.

In this way 278 mg ofN-(2-propenyloxy)-1,2,3,6-tetrahydro-3-pyridinamine of molecular formulaC₈H₁₄N₂O (M=154.21 g) is obtained.

The corresponding yield is 73%.

Stage E

270 mg (1.75 mmoles) of the product obtained in Stage D is dissolvedunder an argon atmosphere in 45 ml of acetonitrile, 760 μl of TEA and105 μl of diphosgene are added.

The reaction medium is left to react for 15 minutes at 0° C., then leftto return to ambient temperature and left to react for another 2 hours.

Then 213 mg of DMAP is added and the medium is left to react overnight.

AcOEt is added, followed by washing with a 10% aqueous solution oftartaric acid and with water.

The organic phase is dried over magnesium sulphate, filtered and thesolvent is evaporated off under reduced pressure.

The crude product obtained is purified on silica, eluting with adichloromethane/acetone mixture 95/5 containing 0.1% TEA.

In this way 36 mg of6-(2-propenyloxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one of molecularformula C₉H₁₂N₂O₂ (M=180.21 g) is recovered.

The corresponding yield is 11%.

Stage F

The operation is carried out in a similar manner to that described inStage E of Example 52 starting from 51 mg (0.27 mmole) of the productobtained in Stage E, 33 μl of acetic acid, 165 mg of Pd[P(C₆H₅)₃]₄ and132 mg of SO₃-pyridine complex.

In this way 29.6 mg of the 1-propenyltriphenylphosphonium salt of6-(sulphooxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one is recovered.

This salt is passed through a column of DOWEX 50WX8 resin in Na⁺ form,followed by eluting with water containing 10% THF.

The product collected is lyophilized in order to obtain 13 mg of theexpected sodium salt, of molecular formula C₆H₇N₂O₅SNa (M=242.19 g).

The corresponding yield is 20%.

MS (negative electrospray) m/z: [M]⁻=219

Example 54 Sodium salt of6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octan-7-one

The operation is carried out as indicated in Stage A of Example 53starting from 12 g (0.061 mole) of 1,1-dimethylethyl3,6-dihydro-3-oxo-1(2H)-pyridinecarboxylate of molecular formulaC₁₀H₁₅NO₃, 9.7 g of O-benzylhydroxylamine hydrochloride and 15 ml ofpyridine.

In this way 19.4 g of 1,1-dimethylethyl3,6-dihydro-3-[(phenylmethoxy)imino]-1(2H)-pyridinecarboxylate ofmolecular formula C₁₇H₂₂N₂O₃ (M=302.38 g) is obtained.

The corresponding yield is quantitative.

Stage B

The operation is carried out as indicated in Stage B of Example 53starting from 14.9 g (0.0496 mole) of the product obtained in Stage A,12 g of sodium cyanoborohydide and 30 ml of boron trifluoride etherate.In this way 8.2 g of a mixture of 2/3 of 1,1-dimethylethyl3,6-dihydro-3-[(phenylmethoxy)amino]-1(2H)-pyridinecarboxylate and 1/3of 1,1-dimethylethyl 3-[(phenylmethoxy)amino]-1-piperidinecarboxylate ofmolecular formula C₁₇H₂₄N₂O₃ (M=304.39 g) is obtained afterpurification.

The corresponding yield is 55%.

Stage C

The operation is carried out as indicated in Stage C of Example 53starting from 9.3 g (0.0306 mole) of a mixture obtained in Stage B and106 ml of a solution of hydrogen chloride in AcOEt at 7 mol/l.

In this way 8.39 g of a mixture of 2/3 ofN-(phenylmethoxy)-1,2,3,6-tetrahydro-3-pyridinamine dihydrochloride and1/3 of N-(phenylmethoxy)-3-piperidinamine dihydrochloride of molecularformula C₁₂H₁₈Cl₂N₂O (M=277.20 g) is obtained.

The corresponding yield is 98%.

Stage D

The operation is carried out as indicated in Stage D of Example 53starting from 8.30 g (0.0299 mole) of the mixture obtained in Stage Cand 30 ml of 2N soda.

In this way 5.95 g of mixture of 2/3 ofN-(phenylmethoxy)-1,2,3,6-tetrahydro-3-pyridinamine and 1/3 ofN-(phenylmethoxy)-3-piperidinamine of molecular formula C₁₂H₁₆N₂O(M=204.27 g) is obtained.

The corresponding yield is 98%.

Stage E

The operation is carried out as indicated in Stage E of Example 53starting from 5.02 g (0.0246 mole) of a mixture obtained in Stage D,2.43 ml of diphosgene, 7.4 ml of TEA and 3 g of DMAP.

In a flask equipped of a magnetic stirrer, 5.020 g (0.0246 mole) of theproduct obtained in Stage D and 1.2 ml of 1,2-dichloroethane areintroduced at 0° C. and under argon.

2.43 g of diphosgene is added.

In this way 2.4 g of6-(phenylmethoxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one of molecularformula C₁₃H₁₄N₂O₂ (M=230.27 g) is recovered after purification. Thecorresponding yield is 42%.

512 mg of 6-(phenylmethoxy)-1,6-diazabicyclo[3.2.1]octan-7-one ofmolecular formula C₁₃H₁₆N₂O₂ (M=232.27 g) is also recovered.

The corresponding yield is 9%.

Stage F

0.128 g (0.551 mmole) of6-(phenylmethoxy)-1,6-diazabicyclo[3.2.1]octan-7-one obtained in Stage Eis dissolved in 1 ml of methanol.

0.035 g of Pd/C catalyst is added and the reaction medium is placedunder a hydrogen atmosphere at normal pressure. At the end of thereaction, the reaction medium is filtered, rinsed with methanol and thesolvent is evaporated off under reduced pressure.

In this way 76 mg of 6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one ofmolecular formula C₆H₁₀N₂O₂ (M=142.16 g) is obtained.

The corresponding yield is quantitative.

Stage G

In a flask placed under an inert atmosphere, 75 mg (0.528 mmole) of theproduct obtained in Stage F is introduced into 2 ml of pyridine.

235 mg of SO₃-pyridine complex is added and the reaction medium is leftto react for 2 hours.

Then a few drops of water are added and the solvent is evaporated offunder reduced pressure.

In this way 361 mg of crude product is obtained which is purified bychromatography on silica eluting with a dichloromethane/ethanol mixture6/4 containing 0.1% by weight of TEA.

In this way 32 mg of the purified triethylammonium salt of6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octan-7-one, of molecular formulaC₁₁H₁₅N₃O₅S (M=301.32 g) is recovered.

The corresponding yield is 17%.

Stage H

31 mg of the product obtained in Stage G is dissolved in 0.5 ml of watercontaining 10% THF.

The solution obtained is passed through a column of DOWEX 50WX8 resin inNa⁺ form, eluting with water containing 10% THF.

The product collected is lyophilized in order to obtain 20 mg of theexpected sodium salt, of molecular formula C₉H₉N₂O₅SNa (M=221 g).

The corresponding yield is 77%.

MS (negative electrospray) m/z: [M−H]⁻=221

Pharmacological Study of the Products of the Invention

I/The compounds of formula (I) and their pharmaceutically acceptablesalts show marked inhibitory activities against the β-lactamases ofvarious bacterial strains and these useful therapeutic properties can bedetermined in vitro on isolated β-lactamases:

A. Preparation of the β-Lactamases Tem-1 and P99

The β-lactamases are isolated from bacterial strains which are resistantto penicillins and cephalosporins (Tem1 and P99 are produced by E. coli250HT21 and E. Cloacae 293HT6 respectively).

The bacteria are cultured at 37° C. in heart-brain culture at 37 g/l(DIFCO). They are harvested at the end of the exponential phase, cooleddown and centrifuged. The bacterial pellets are taken up in sodiumphosphate buffer 50 mM, pH 7.0 and centrifuged again. The bacteria aretaken up in two volumes of the same buffer and lyzed using aFrench-Press maintained at 4° C. After centrifugation for 1 hour at100,000 g, at 4° C., the supernatants containing the soluble fraction ofthe bacterial extracts are recovered and frozen at −80° C.

B. Determination of the β-Lactamases Activity

The method uses Nitrocefin (OXOID), cephalosporin chromogene assubstrate, of which the product hydrolyzed by the B-lactamases is redand absorbs at 485 nm. Beta-lactamase activity is determined kineticallyby measurement of the absorbance variation at 485 nm resulting from thehydrolysis of the substrate using a plate spectrophotometer (Spectra MaxMore Molecular Devices). The experiments are carried out at 37° C. Thequantity of enzyme was normalized and the measurements are carried outat initial speed.

C. Determination of Compound Inhibitory Activity on Lactamases

Two measurements were carried out, without preincubation and withpreincubation of the enzyme and inhibitor (5 nm), in order to test theirreversibility of the reaction. The products are tested at 6 or 8concentrations in duplicate. The reaction mixture contains 100 μM ofNitrocefin and 50 mM sodium phosphate buffer pH7.0.

D. Calculations of the IC50

The hydrolysis speeds are measured with and without inhibitor. Theconcentration of inhibitor which inhibits by 50% the hydrolysis reactionof Nitrocefin by the enzyme (CI50) is determined. The data is processedusing GraFit software (Erathycus Software).

EXAMPLE n^(o) IC₅₀ nM/TEM1 IC₅₀ nM/P99  1a 700 (>10 000)  2 462 —  2a6730 —  3 590 9800  3a 4400 —  3b 2010 —  4 2710 —  5 1010 —  7 650 250 7a 55 17  8 1400 62  9 8500 630 10 0.26 1.50 14 6400 — 19^(e) 11 — 19f110 — 19g 29 — 22 5100 — 25 28 600 26a 115 1850 26b 4900 26c 1100 700028b 9.5 12 28c 29 1100 28d 1.3 390 28^(e) 52 — 29b 460 4200 29c 450 —29d 9500 2000 29^(e) 4200 6300 29f 5200 — 30 3500 — 31 5700 — 33 17 33034 27 32 35 53 56 36 23 110 37 29 160 38 35 77 39 31 50 40 51 96 41 14120 42 25 70 43 31 76 44 59 100 45 12 60 46 26 70 47 18 43 48 15 120 498.2 98 50 18 150 52 11 4600 53 15 5900 54 3100 —

II/Inhibition of the β-lactamases shown above potentiates the antibioticactivity of β-lactamine type antibiotics, thus leading to a synergiceffect, as the following results also show, which express the minimalinhibitory concentration in vitro (MIC in μg/ml), vis-à-vis a certainnumber of pathogenic microorganisms, of combinations of cefotaxime andpiperacillin with compounds of formula (I) at a concentration of 5 mg/l.

The operation is carried out as follows, using the so-calledmicro-dilution in liquid medium method.

A series of concentrations of the β-lactamine is prepared in thepresence of a constant concentration (5 mg/l) of the product to bestudied, each is then seeded with various bacterial strains.

After incubation for 24 hours in a heating chamber at 37° C., the growthinhibition is evaluated by the absence of any bacterial development,which allows the minimal inhibitory concentrations (MIC) for each strainto be determined, expressed in milligrams/l.

The following results were obtained:

Trial Cefo- Wt. Ex. N^(o) Strain Phenotype taxime Ex. 35 33 1 011GO66 S.aureus PeniR 1.2 1.2 2.5 2 250HT21 E. coli Tem1 <=0.04 <=0.04 <=0.04 3250HT22 E. coli Tem2 <=0.04 <=0.04 <=0.04 4 250CF1 E. coli Tem3 >40 0.15<=0.04 5 250SJ1 E. coli Tem7 0.08 <=0.04 <=0.04 6 250HT26 E. coli SHV10.6 <=0.04 <=0.04 7 250BE1 E. coli SHV4 40 0.08 <=0.04 8 250HT23 E. coliClass D 0.08 <=0.04 <=0.04 9 293HT6 E. cloacae Class C >40 0.6 <=0.04 10301HT6 Serratia Serratia 0.3 <=0.04 0.08 11 391HT7 P. aeruginosa PSE 4020 20 12 391HT8 P. aeruginosa PSE 40 20 20 Trial N^(o) Ex. 34 Ex. 36 Ex.37 Ex. 38 Ex. 39 Ex. 40 Ex. 41 1 1.2 1.2 1.2 1.2 2.5 2.5 2.5 2 <=0.04<=0.04 <=0.04 <=0.04 <=0.04 <=0.04 <=0.04 3 <=0.04 <=0.04 <=0.04 <=0.04<=0.04 <=0.04 <=0.04 4 0.15 0.3 0.3 0.3 0.08 0.6 2.5 5 <=0.04 <=0.040.08 <=0.04 <=0.04 0.15 0.6 6 <=0.04 <=0.04 <=0.04 <=0.04 <=0.04 <=0.04<=0.04 7 0.08 0.3 0.6 0.3 0.08 1.2 >40 8 <=0.04 <=0.04 <=0.04 <=0.04<=0.04 <=0.04 0.08 9 0.6 20 >40 20 1.2 >40 >40 10 <=0.04 0.08 0.08 0.08<=0.04 0.08 0.08 11 20 40 20 40 20 >40 >40 12 20 40 20 40 20 >40 >40Trial N^(o) Ex. 42 Ex. 43 Ex. 44 Ex. 48 Ex. 49 1 2.5 2.5 2.5 2.5 2.5 2<=0.04 <=0.04 <=0.04 <=0.04 <=0.04 3 <=0.04 <=0.04 0.15 <=0.04 <=0.04 4<=0.04 0.08 0.3 <=0.04 0.08 5 <=0.04 <=0.04 0.08 <=0.04 <=0.04 6 <=0.04<=0.04 0.08 <=0.04 <=0.04 7 <=0.04 <=0.04 0.6 0.08 <=0.04 8 <=0.04<=0.04 <=0.04 <=0.04 <=0.04 9 0.3 0.08 >40 0.15 Nd 10  <=0.04 <=0.04 10<=0.04 Nd 11  20 20 >40 40 Nd 12  20 20 >40 >40 nd Trial Wt. N^(o)Strain Phenotype Piperacillin Ex. 35 Ex. 33 Ex. 34 1 011GO66 peniR 101.2 1.2 0.6 2 250HT21 Tem1 >40 2.5 0.3 5 3 250HT22 Tem2 >40 20 0.6 >40 4250CF1 Tem3 >40 10 1.2 >40 5 250SJ1 Tem7 >40 5 0.6 >40 6 250HT26SHV1 >40 5 1.2 20 7 250BE1 SHV4 >40 20 1.2 >40 8 250HT23 Class D >40 102.5 40 9 293HT6 Class >40 5 0.6 10 10 301HT6 Serratia 5 1.2 0.6 1.2 11391HT7 PSE >40 >40 >40 >40 12 391HT8 PSE >40 >40 10 >40 Trial N^(o) Ex.36 Ex. 37 Ex. 38 Ex. 39 Ex. 40 Ex. 41 Ex. 42 1 1.2 0.6 0.6 1.2 0.6 1.21.2 2 5 2.5 5 2.5 10 40 1.2 3 40 >40 >40 20 >40 >40 2.5 4 40 40 2040 >40 >40 2.5 5 20 >40 >40 >40 >40 >40 2.5 6 20 20 40 40 >40 >40 57 >40 >40 >40 20 >40 >40 5 8 20 40 >40 10 20 40 5 9 20 40 40 20 >40 >401.2 10 1.2 2.5 1.2 0.6 1.2 1.2 0.6 11 >40 >40 >40 >40 >40 >40 >4012 >40 >40 >40 >40 >40 >40 >40 Trial Ex. Ex. Ex. Ex. N^(o) Ex. 43 Ex. 44Ex. 45 Ex. 46 47 48 49 50 1 1.2 2.5 1.2 0.6 0.6 1.2 1.2 1.2 2 1.2 5 1.25 2.5 0.6 1.2 2.5 3 1.2 >40 2.5 >40 20 2.5 2.5 40 4 2.5 40 2.5 2.5 5 1.21.2 5 5 5 >40 2.5 >40 40 1.2 5 40 6 5 >40 2.5 >40 2.5 1.2 20 40 7 10 >405 40 5 2.5 20 40 8 2.5 40 10 10 10 5 10 40 9 2.5 >40 1.2 >40 >40 2.510 >40 10 0.6 >40 0.6 1.2 1.2 0.6 nd 0.6 11 >40 >40 >40 >40 >40 >40nd >40 12 >40 >40 >40 >40 >40 >40 nd >40

Example of a Pharmaceutical Composition:

1/A pharmaceutical composition (lyophilisates) for injection wasprepared containing

on the one hand: compound of Example 35 500 mg on the other hand:Cefotaxime  1 g Sterile aqueous excipient s.q.f.  5 cm³

The two active ingredients can, if desired, be introduced separatelyinto two distinct ampoules or flasks.

2/A pharmaceutical composition (lyophilisates) for injection wasprepared containing

on the one hand: compound of Example 33 250 mg on the other hand:Cefpirone  1 g Excipient sterile aqueous s.q.f.  5 cm³

The two active ingredients can, if desired, be introduced separatelyinto two distinct ampoules or flasks.

1-18. (canceled)
 19. A pharmaceutical composition comprising (a) acompound, or a pharmaceutically acceptable salt thereof, of formula (I):

wherein R₁ is hydrogen, COOH, CN, COOR, CONR₆R₇, (CH₂)_(n′)R₅ orC(═NR₆)NHR₇; R is selected from the group consisting of alkyl containing1 to 6 carbon atoms optionally substituted by a pyridyl or carbamoylradical, —CH₂-alkenyl containing 3 to 9 carbon atoms, aryl containing 6to 10 carbon atoms and aralkyl containing 7 to 11 carbon atoms, whereinthe nucleus of said aryl or aralkyl is optionally substituted by OH,NH₂, NO₂, alkyl containing 1 to 6 carbon atoms, alkoxy containing 1 to 6carbon atoms or by one or more halogen atoms; R₆ and R₇ are identical ordifferent and are independently selected from the group consisting ofhydrogen, alkyl containing 1 to 6 carbon atoms, aryl containing 6 to 10carbon atoms and aralkyl containing 7 to 11 carbon atoms optionallysubstituted by a carbamoyl, ureido or dimethylamino radical, and alkylcontaining 1 to 6 carbon atoms substituted by a pyridyl radical; n′ is 1or 2; R₅ is selected from the group consisting of COOH, CN, OH, NH₂,CO—NR₆R₇, COOR, OR, OCHO, OCOR, OCOOR, OCONHR, OCONH₂, NHR, NHCOH,NHCOR, NHSO₂R, NH—COORU, NH—CO—NHR and NHCONH₂ wherein R, R₆ and R₇ areas defined above; R₂ is hydrogen or (CH₂)_(n′1)R₅ wherein n′₁ is 0, 1 or2, and R₅ is as defined above; R₃ is hydrogen or alkyl containing 1 to 6carbon atoms; A is a bond between the two carbons which carry R₁ and R₂,

group wherein R₄ is hydrogen or (CH₂)_(n′1)R₅ and n′₁ and R₅ are asdefined above, and the dotted line is an optional bond with one of thetwo carbons which carry R₁ and R₂; n is 1 or 2; X is a divalent —C(O)-B-group linked to the nitrogen atom by the carbon atom wherein B is adivalent —O—(CH₂)_(n′)— group linked to the carbonyl by the oxygen atom,a divalent —NR₈—(CH₂)_(n′1)— or —NR₈—O— group linked to the carbonyl bythe nitrogen atom, n″ is 0 or 1, and wherein B is —NR₈—(CH₂)_(n″)—, R₈is selected from the group consisting of hydrogen, OH, R, OR, Y, OY, Y₁,OY₁, Y₂, OY₂, Y₃, OCH₂CH₂SO_(m)R, OSiR_(a)R_(b)R_(c) andSiR_(a)R_(b)R_(c) and wherein B is —NR₈—O—, R₈ is selected from thegroup consisting of hydrogen, R, Y, Y₁, Y₂, Y₃ and SiR_(a)R_(b)R_(c),wherein R_(a), R_(b) and R_(c) is each independently a linear orbranched alkyl containing 1 to 6 carbon atoms or aryl containing 6 to 10carbon atoms, R is as defined above and m is 0, 1 or 2; Y is selectedfrom the group consisting of COH, COR, COOR, CONH₂, CONHR, CONHOH,CONHSO₂R, CH₂COOH, CH₂COOR, CH₂CONHOH, CH₂CONHCN, CH₂tetrazole,protected CH₂tetrazole, CH₂SO₃H, CH₂SO₂R, CH₂PO(OR)₂, CH₂PO(OR)(OH),CH₂PO(R)(OH) and CH₂PO(OH)₂; Y₁ is selected from the group consisting ofSO₂R, SO₂NHCOH, SO₂NHCOR, SO₂NHCOOR, SO₂NHCONHR, SO₂NHCONH₂ and SO₃H; Y₂is selected from the group consisting of PO(OH)₂, PO(OR)₂, PO(OH)(OR)and PO(OH)(R); Y₃ is selected from the group consisting of tetrazole,tetrazole substituted by R, squarate, NH or NR-tetrazole, NH orNR-tetrazole substituted by R, NHSO₂R and NRSO₂R wherein R is as definedabove; and R₁, R₂ and R₃ are not simultaneously hydrogen when n is 1, Ais

wherein R₄ is hydrogen and X is —C(O)—O—(CH₂)_(n″) wherein n″ is 0 or 1,or X is —CO—NR₈—(CH₂)_(n″) wherein n″ is 1 and R₈ is isopropyl, or X is—CO—NR₈—(CH₂)_(n″) wherein n″ is 0 and R₈ is hydrogen or phenyl; and (b)a therapeutically effective amount of a β-lactamine antibiotic.
 20. Apharmaceutical composition according to claim 19, wherein saidβ-lactamine antibiotic comprises an antibiotic selected from the groupconsisting of: penams, penems, carbapenems, cephems, carbacephems,oxacephems, cephamycims, monobactams, and combinations andpharmaceutically acceptable salts thereof.
 21. A pharmaceuticalcomposition according to claim 19, wherein said β-lactamine antibioticcomprises a cephalosporin antibiotic selected from the group consistingof: cephalothin, cephaloridine, cefaclor, cefadroxil, cefamandole,cefazolin, cephalexin, cephradine, ceftizoxime, cefoxitin, cephacetrile,cefotiam, cefotaxime, cefsulodin, cefoperazone, ceftizoxime,cefmenoxime, cefmetazole, cephaloglycin, cefonicid, cefodizime,cefpirome, ceftazidime, ceftriaxone, cefpiramide, cefbuperazone,cefozopran, cefepime, cefoselis, cefluprenam, cefuzonam, cefpimizole,cefclidin, cefixime, ceftibuten, cefdinir, cefpodoxime axetil,cefpodoxime proxetil, cefteram pivoxil, cefetamet pivoxil, cefcapenepivoxil, cefditoren pivoxil, cefuroxime, cefuroxime axetil,loracarbacef, latamoxef, and combinations and pharmaceuticallyacceptable salts thereof.
 22. A pharmaceutical composition according toclaim 21, wherein said β-lactamine antibiotic comprises ceftazidime. 23.The pharmaceutical composition according to claim 19, wherein n is 1, R₃is hydrogen, R₁ is hydrogen, COOR or CONR₆R₇ wherein R, R₆ and R₇ are asdefined in claim 19, and X is —C(O)-B- wherein B is —O—(CH₂)_(n″)—or—NR₈—(CH₂)_(n″)— wherein n″ is 0 and R₈ is as defined in claim
 19. 24.The pharmaceutical composition according to claim 19, wherein R₈ is Y,Y₁ or OY₁.
 25. The pharmaceutical composition according to claim 19,wherein R₂ and R₄ are each hydrogen, and B is —NR₈—(CH₂)_(n″)— whereinn″ is 0 and R₈ is OY₁.
 26. The pharmaceutical composition according toclaim 25, wherein R₃ is hydrogen, and R₁ is hydrogen, COOR or CONR₆R₇.27. The pharmaceutical composition according to claim 19, wherein thecompound of Formula (I) is selected from the group consisting of:cis-7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-4-propanoic acid, transdiphenylmethyl 7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetate, cisdiphenylmethyl 7-oxo-6-oxa-1-azabicyclo[3.2.1]octan-4-acetate, transphenylmethyl3-benzoyl-2-oxo-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate, transphenylmethyl2-oxo-3-(sulphooxy)-1,3-diazabicyclo[2.2.1]heptane-6-carboxylate,6-[[(4-methylphenyl)sulphonyl]oxy]-1,6-diazabicyclo[3.2.1]octan-7-one,6-[(methylsulphonyl)oxy]-1,6-diazabicyclo[3.2.1]octan-7-one,6-[(4-nitrophenyl)sulphonyl]oxy]-1,6-diazabicyclo[3.2.1]octan-7-one,trans diphenylmethyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-2-carboxylate, trans(4-nitrophenyl)methyl7-oxo-6-oxa-1-azabicyclo[3.2.1]octane-2-carboxylate,trans-7-oxo-6-oxa-1-azabicyclo[3.2.1.]octane-2 carboxylic acid, transphenylmethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate, transphenylmethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate, transphenylmethyl7-oxo-6-[(phenylsulphonyl)oxy]-1,6-diaza-bicyclo[3.2.1]octane-2-carboxylate,trans phenylmethyl7-oxo-6-[(2-thienylsulphonyl)oxy]-1,6-diazabicyclo[3-.2.1]octane-2-carboxylattrans-6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid,trans methyl6-benzoyl-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,trans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-7-oxo-N-(phenylmethyl)-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-7-oxo-N-(2-pyridinylmethyl)-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-7-oxo-N-[2-(3-pyridinyl)ethyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-7-oxo-N-[2-(4-pyridinyl)ethyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-7-oxo-N-[2-(2-pyridinyl)ethyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-N-[3-(aminocarbonyl)phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.-2.1] octane-2-carboxamide,trans-N-[4-(dimethylamino)phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-N-[3-(dimethylamino)phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-7-oxo-N-[(4-pyridinyl)methyl]-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-7-oxo-N-(3-pyridinylmethyl)-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-N-(1-amino-1-oxo-3-phenyl-2-propyl)-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-N-(2-amino-2-oxoethyl)-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-N-[3-[(aminocarbonyl)amino]phenyl]-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-N-(2-amino-2-oxo-1-phenylethyl)-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide,trans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate of2-amino-2-oxoethyl, trans 2-(4-pyridinyl)ethyl7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1] octane-2-carboxylate, trans2-(2-pyridinyl)ethyl 7-oxo-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxylate,6-(sulphooxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one, and3-methoxy-6-(sulphooxy)-1,6-diazabicyclo[3.2.1]oct-3-in-7-one.
 28. Thepharmaceutical composition of claim 27, wherein said β-lactamineantibiotic is selected from the group consisting of: penams, penems,carbapenems, cephems, carbacephems, oxacephems, cephamycims,monobactams, and combinations and pharmaceutically acceptable saltsthereof.
 29. The pharmaceutical composition of claim 27, wherein saidβ-lactamine antibiotic is selected from the group consisting of:amoxicillin, ampicillin, azlocillin, mezlocillin, apalcillin,hetacillin, bacampicillin, carbenicillin, sulbenicillin, ticarcillin,piperacillin, azlocillin, mecillinam, pivmecillinam, methicillin,ciclacillin, talampicillin, aspoxicillin, oxacillin, cloxacillin,dicloxacillin, flucloxacillin, nafcillin, pivampicillin, cephalothin,cephaloridine, cefaclor, cefadroxil, cefamandole, cefazolin, cephalexin,cephradine, ceftizoxime, cefoxitin, cephacetrile, cefotiam, cefotaxime,cefsulodin, cefoperazone, ceftizoxime, cefmenoxime, cefmetazole,cephaloglycin, cefonicid, cefodizime, cefpirome, ceftazidime,ceftriaxone, cefpiramide, cefbuperazone, cefozopran, cefepime,cefoselis, cefluprenam, cefuzonam, cefpimizole, cefclidin, cefixime,ceftibuten, cefdinir, cefpodoxime axetil, cefpodoxime proxetil, cefterampivoxil, cefetamet pivoxil, cefcapene pivoxil, cefditoren pivoxil,cefuroxime, cefuroxime axetil, loracarbacef, latamoxef, imipenem,meropenem, biapenem, panipenem, aztreonam, carumonam, and combinationsand pharmaceutically acceptable salts thereof.
 30. The pharmaceuticalcomposition of claim 27, wherein said β-lactamine antibiotic is selectedfrom the group consisting of: cephalothin, cephaloridine, cefaclor,cefadroxil, cefamandole, cefazolin, cephalexin, cephradine, ceftizoxime,cefoxitin, cephacetrile, cefotiam, cefotaxime, cefsulodin, cefoperazone,ceftizoxime, cefmenoxime, cefmetazole, cephaloglycin, cefonicid,cefodizime, cefpirome, ceftazidime, ceftriaxone, cefpiramide,cefbuperazone, cefozopran, cefepime, cefoselis, cefluprenam, cefuzonam,cefpimizole, cefclidin, cefixime, ceftibuten, cefdinir, cefpodoximeaxetil, cefpodoxime proxetil, cefteram pivoxil, cefetamet pivoxil,cefcapene pivoxil, cefditoren pivoxil, cefuroxime, cefuroxime axetil,loracarbacef, latamoxef, and combinations and pharmaceuticallyacceptable salts thereof.
 31. The pharmaceutical composition accordingto claim 19, wherein said compound of Formula (I) comprisestrans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo-[3.2.1]-octane-2-carboxamideor a pharmaceutically acceptable salt thereof.
 32. The pharmaceuticalcomposition according to claim 19, wherein said compound of Formula (I)comprises a sodium salt oftrans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo-[3.2.1]-octane-2-carboxamide.33. The pharmaceutical composition of claim 19, wherein said compound ofFormula (I) comprisestrans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo-[3.2.1]-octane-2-carboxamideor a pharmaceutically acceptable salt thereof and said β-lactamineantibiotic comprises ceftazidime.
 34. The pharmaceutical composition ofclaim 19, wherein said compound of Formula (I) comprises a sodium saltoftrans-7-oxo-6-(sulphooxy)-1,6-diazabicyclo-[3.2.1]-octane-2-carboxamideand said β-lactamine antibiotic comprises ceftazidime.